Oxadiazole Diaryl Compounds

ABSTRACT

The invention relates to compounds of formula (I): 
     
       
         
         
             
             
         
       
     
     wherein R 1 , R 2 , R a , R b , R c  and W, have the meanings given in claim  16 . The compounds are useful e.g. in the treatment of autoimmune disorders, such as multiple sclerosis.

The present invention relates to oxadiazole diaryl compounds, their useas medicaments and their use for treating multiple sclerosis and otherdiseases.

In particular, the invention relates to compounds of formula (I):

wherein

-   -   R¹, R² denote H, Hal, CF₃, OCF₃, CN, or NO₂,    -   W denotes CH or N,    -   R^(a) is Ar, Het, cycloalkyl having 3-7 atoms, A or NA₂,    -   R^(b) is H, A, Hal, CF₃, OCF₃, OR³, CN, NO₂, (CH₂)_(n)N(R³)₂,        OA, especially OCH₃, (CH₂)_(n)SO₂N(R³)₂, (CH₂)_(n)NR³SO₂A,        (CH₂)_(n)N(SO₂A)₂, NR³CON(R³)₂, NR³COA or (CH₂)_(n)SO₂R³,    -   R^(c) denotes A, COA, CSA, CODA, CSOA, CON(R³)₂ or CSN(R³)₂    -   A is branched or linear alkyl having 1 to 12 C-atoms, wherein        one or more, preferably all H-atoms may be replaced by Hal,        especially F, or 1 to 7H-atoms may be replaced by OR³, CN or        N(R³)₂ and wherein one or more, preferably 1 to 7 non-adjacent        CH₂-groups may be replaced by O, NR³ or S and/or by —CH═CH— or        —C≡C— groups, or denotes cycloalkyl or cycloalkylalkylen having        3-7 ring C atoms    -   Hal is F, Cl, Br or I,    -   Ar denotes a monocyclic or bicyclic, unsaturated or aromatic        carbocyclic ring having 6 to 14 carbon atoms which may be        unsubstituted or monosubstituted, disubstituted or        trisubstituted by Hal, A, CH₂OA, CH₂N(R³)₂, OR³, N(R³)₂, NO₂,        N(SO₂Me)₂, CN, COOR³, CF₃, OCF₃, CON(R³)₂, NR³COA, NR³CON(R³)₂,        NR³SO₂A, COR³, SO₂N(R³)₂, SOA or SO₂A, phenyl, pyridyl        —[C(R³)₂]_(n)—COOR³ and/or —O[C(R³)₂]_(n)—CON(R³)₂,    -   Het denotes a monocyclic or bicyclic, saturated, unsaturated or        aromatic heterocyclic ring having 1 N and/or 1 to 3 O and/or S        atoms which may be unsubstituted or monosubstituted,        disubstituted or trisubstituted by Hal, A, —[C(R³)₂]_(n)—Ar,        —[C(R³)₂]_(n)-cycloalkyl, CH₂OA, CH₂N(R³)₂, OR³, CF₃, OCF₃,        N(R³)₂, N(SO₂Me)₂, NR³CON(R³)₂, NO₂, CN, —[C(R³)₂]_(n)—COOR³,        —[C(R³)₂]_(n)—CON(R³)₂, NR³COA, NR³SO₂A, COR³, SO₂N(R³)₂, SOA,        phenyl, pyridyl and/or SO₂A,    -   R³ is H or A    -   and    -   n is 0, 1, 2, 3, 4, 5, 6, 7 or 8,        and pharmaceutically acceptable derivatives, solvates,        tautomers, salts and stereoisomers thereof, including mixtures        thereof in all ratios.

The compounds of formula (I) are preferably binding on receptors forsphingosine 1-phosphate (S1P). S1P is a bioactive sphingolipidmetabolite that is secreted by hematopoietic cells and stored andreleased from activated platelets. It acts as an agonist on a family ofG protein-coupled receptors (GPCR). Five sphingosine 1-phosphatereceptors have been identified (S1P₁, S1P₂, S1P₃, S1P₄, and S1P₅, alsoknown as endothelial differentiation genes, which are Edg1, Edg5, Edg3,Edg6 and Edg8 respectively), that have widespread cellular and tissuedistribution and are well conserved in human and rodent species.

S1P is involved in a number of cellular functions such as survival,proliferation and immunological responses. The compounds of the presentinvention are preferably acting as S1P₁/Edg1 receptor agonists and thushave immunosuppressive activities by modulating leukocyte trafficking,sequestering lymphocytes in secondary lymphoid tissues, and interferingwith cell-cell interactions required for an efficient immune response.The invention is also directed to pharmaceutical compositions containingsuch compounds and methods of treatment or prevention.

FTY720 or fingolimod, a non selective S1P₁ agonist, exertsimmunosuppressive activity and shows therapeutic effects in thetreatment of relapsing-remitting multiple sclerosis. Numerouspublications have been already published using this compound: Cyster J GAnnu Rev Immunol 23:127-59, 2005, Rosen H Nat Rev Immunol 5:560-570,2005, Rosen H Trends Immunol 28:102-107, 2007, Yopp A C Clin Transplant20:788-795, 2006, Kappos L N Engl J Med 355:1124-1140, 2006, Massberg SN Engl J Med 355:1088-1089, 2006.

Immunosuppressive agents are further useful in a wide variety ofautoimmune and chronic inflammatory diseases, including systemic lupuserythematosus, chronic rheumatoid arthritis, type I diabetes mellitus,inflammatory bowel diseases, biliary cirrhosis, uveitis and otherdisorders such as Crohn's diseases, ulcerative colitis, bullouspemphigoid, sarcoidosis, psoriasis, autoimmune myositis, Wegener'sgranulomatosis, ichthyosis, Graves opthalmopathy, atopic dermatitis andasthma. They are also useful as part of chemotherapeutic regimens forthe treatment of cancers, lymphomas and leukemias.

It has been found that the compounds of the present invention areselective S1P₁ agonists with improved pharmacological and/or otherproperties.

The present invention uses compounds of Formula (I) and pharmaceuticallyusable derivatives, salts, tautomers, solvates and stereoisomersthereof, including mixtures thereof in all ratios, for the preparationof a medicament for the treatment and/or prophylaxis of diseases inwhich the inhibition, activation, regulation, and/or modulation of S1P₁receptor signal transduction plays a role.

Thus, the present invention preferably comprises compounds which areagonists of the S1P₁/Edg1 receptor, especially having selectivity overthe S1P₃/Edg3 receptor. An S1P₁/Edg1 receptor selective agonist hasadvantages over current therapies and extends the therapeutic window oflymphocyte sequestration agents, allowing better tolerability withhigher dosing and thus improving efficacy.

The invention further relates to the manufacture of a medicament for theimprovement of vascular function, either alone or in combination withother active compounds or therapies.

The oxadiazole diaryl compounds according to Formula (I) may be preparedfrom readily available starting materials using the following generalmethods and procedures. It will be appreciated that where typical orpreferred experimental conditions (i.e. reaction temperatures, time,moles of reagents, solvents etc.) are given, other experimentalconditions can also be used unless otherwise stated. Optimum reactionconditions may vary with the particular reactants or solvents used, butsuch conditions can be determined by the person skilled in the art,using routine optimisation procedures.

The following abbreviations refer respectively to the definitions below:

aq. (aqueous), h (hour), g (gram), L (liter), mg (milligram), MHz(Megahertz), μM (micromolar) min. (minute), mm (millimeter), mmol(millimole), mM (millimolar), m.p. (melting point), eq. (equivalent), mL(milliliter), μL (microliter), ACN (acetonitrile), BINAP(2,2′-bis(disphenylphosphino)-1,1′-binaphthalene, BOC(tert-butoxy-carbonyl), CBZ (carbobenzoxy), CDCl₃ (deuteratedchloroform), CD₃OD (deuterated methanol), CH₃CN (acetonitrile), conc.(concentrated), c-hex (cyclohexane), DCC (dicyclohexyl carbodiimide),DCE (dichloroethane), DCM (dichloromethane), DIC (diisopropylcarbodiimide), DIEA (diisopropylethyl-amine), DMF (dimethylformamide),DMSO (dimethylsulfoxide), DMSO-d₆ (deuterated dimethylsulfoxide), EDC(1-(3-dimethyl-amino-propyl)-3-ethylcarbodiimide), ESI (Electro-sprayionization), Ethyl acetate (ethyl acetate), Et₂O (diethyl ether), EtOH(ethanol), FMOC (fluorenylmethyloxycarbonyl), HATU(dimethylamino-([1,2,3]triazolo[4,5-b]pyridin-3-yloxy)-methylene]-dimethyl-ammoniumhexafluorophosphate), HPLC (High Performance Liquid Chromatography),i-PrOH (2-propanol), K₂CO₃ (potassium carbonate), LC (LiquidChromatography), MeOH (methanol), MgSO₄ (magnesium sulfate), MS (massspectrometry), MTBE (Methyl tert-butyl ether), Mtr.(4-Methoxy-2,3,6-trimethylbenzensulfonyl), MW (microwave), NaHCO₃(sodium bicarbonate), NaBH₄ (sodium borohydride), NMM (N-methylmorpholine), NMR (Nuclear Magnetic Resonance), Pd(PPh₃)₄(tetrakis(triphenylphosphine)palladium), POA (phenoxyacetate), PyBOP®(benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate), RT (room temperature), Rt (retention time), sat.(saturated), SPE (solid phase extraction), TBTU(2-(1-H-benzotriazole-1-yl)-1,1,3,3-tetramethyluromium tetrafluoroborate), TEA (triethylamine), TFA (trifluoroacetic acid), THF(tetrahydrofuran), TLC (Thin Layer Chromatography), UV (Ultraviolet).

Depending on the nature of R^(a), R^(b), R^(c), R¹, R² and W differentsynthetic strategies may be selected for the synthesis of compounds ofFormula (I). In the process illustrated in the following schemes R^(a),R^(b), R^(c), R¹, R² and W are as above-defined in the description.

In general, the oxadiazole diaryl compounds according to Formula (I) ofthis invention may be prepared from readily available startingmaterials. If such starting materials are not commercially availablethey may be prepared by standard synthetic techniques. The followinggeneral methods and procedures described hereinafter in the examples maybe employed to prepare compounds of Formula (I).

Generally, compounds of Formula (I) can be prepared by coupling an arylamidoxime of Formula (III) and an aryl carboxylic acid of Formula (IV)(or an activated derivative thereof, such as formula II) and bydehydrating the resulting intermediate, wherein R^(a), R^(b), R^(c), R¹,R² and W are defined as above, as outlined in Scheme 1. Generalprotocols for such couplings and dehydrations are given below in theExamples, using conditions and methods well known to those skilled inthe art to prepare an oxadiazole from an aryl carboxylic acid and anaryl amidoxime, by forming an acid derivative such as an acid chlorideby reacting the carboxylic acid with oxalyl chloride, thionyl chlorideor trichloroacetonitrile and a suitable phosphine (e.g. Polymersupported triphenyl phosphine) in the presence or absence of bases suchas TEA, DIEA, NMM in a suitable solvent such as DCM, THF or DMF, at atemperature rising from 20° C. to 200° C., for a few hours, e.g. onehour to 24 h. The obtained acid chloride may be coupled to an amidoximeof Formula (III) and the resulting intermediate dehydrated in thepresence or absence of bases such as TEA, DIEA, NMM or pyridine in asuitable solvent such as DCM, THF or toluene at a temperature risingfrom about 20° C. to about 200° C., preferably 150° C. for a timeranging from about 10 minutes to about 24 h, preferably 30 minutes.Alternatively, coupling agents, such as but not limited topolymer-supported 1-alkyl-2-chloropyridinium salt (polymer-supportedMukaiyama's reagent), 1-methyl-2-chloropyridinium iodide (Mukaiyama'sreagent), EDC or HATU in the presence or absence of bases such as TEA,DIEA, NMM in a suitable solvent such as DCM, THF or DMF, at atemperature rising from about 20° C. to about 50° C., preferably at roomtemperature, for a few hours, e.g. one hour to 24 h, may be used.

Compounds of Formula (Ia) can be obtained from Compounds of Formula (Ib)as outlined in Scheme 2 by the reduction of the nitro to an amine, usinghydrogen (which can be generated in situ from ammonium formate or formicacid) in the presence of a metal catalyst such as Pd/C or Raney nickel,or using reducing agents such as iron or tin(II) chloride, or usingmetal hydrides such as lithium aluminum hydride, in a suitable solventsuch as a lower alcohol, water, THF, dioxane or ether, or a mixturethereof, at a temperature ranging from about −100° C. to about 100° C.,preferably from 0° C. to 80° C., for a few hours.

Compounds of Formula (Ic) and/or (Id), can be obtained from compounds ofFormula (Ie) as outlined in Scheme 3, by sulfonylation of the aminogroup, using alkylsulfonylchlorides. This reaction can be done in theabsence or the presence of a base such as TEA, DIEA or pyridine, in asuitable solvent such as DCM, DCE, THF, dioxane, DMF or DMA or a mixturethereof. This reaction is preferrably performed at a temperature rangingfrom about 0° C. to about 100° C., preferably from 0° C. to 40° C., fora few hours.

Compounds of Formula (If) can be obtained from compounds of Formula (Ih)as outlined in Scheme 4 by transforming the hydroxyl moiety into aleaving group such as, but not limited to, alkyl- or aryl-sulfonates orhalogens (preferably Cl, Br and I). The obtained intermediate (Ig) isthen reacted with an amine of Formula HN(R³)₂ in the absence or thepresence of a base such as TEA, DIEA or pyridine, in a suitable solventsuch as DCM, DCE, THF, dioxane DMF or DMA or a mixture thereof. Thisreaction is preferrably performed at a temperature ranging from about 0°C. to about 100° C., preferably from 0° C. to 40° C., for a few hours,preferably from 1 to 5 hours.

Compounds of Formula (II) can be obtained from compounds of Formula (Ih)by one step oxidation using reagents such as KMnO₄ or K₂Cr₂O₇. Compoundsof Formula (Ii) can also be obtained by a two-step procedure as outlinedin scheme 5. The first step is the transformation of the —CH₂OH moietyinto an aldehyde (—CHO) using conventional conditions such as, but notlimited to, MnO₂ or Swern oxidation. The second step consists inreacting the intermediate (ID with an oxidant such as sodium chlorite.This reaction can be performed in the presence or the absence of a basesuch as sodium dihydrogenphosphate, in a suitable solvent such as water,THF, dioxane or a mixture thereof. This reaction is preferrablyperformed at a temperature ranging from about −20° C. to about 60° C.,preferably from 0° C. to 40° C., for a few hours, preferably from 10 to40 hours.

The compounds of Formula (III), wherein R¹, R² and R^(c) are defined asabove, can be obtained as outlined in Scheme 6 by reacting acommercially available aryl nitrile derivative with hydroxylamine in asuitable solvent such as water, MeOH, EtOH or a mixture thereof,preferably EtOH at a temperature ranging from about 20° C. to about 100°C., preferably 20° C. to 60° C. for a few hours.

The method for preparing the compounds of Formula (III) selected below:

-   N′-hydroxy-2-methoxybenzenecarboximidamide-   N′-hydroxy-2-(trifluoromethoxy)benzenecarboximidamide-   2-ethoxy-N′-hydroxybenzenecarboximidamide-   5-fluoro-N′-hydroxy-2-methoxybenzenecarboximidamide    is more particularly described in the Examples.

The compounds of Formula (IV), wherein R^(a) and R^(b) are defined asabove, can be obtained as outlined in Scheme 7 when R^(a) is a tertiaryamino group. The first step consists of the reaction of a commerciallyavailable secondary amine (R^(a)—H) with a commercially available4-fluorobenzoic ester derivative in the presence or absence of basessuch as TEA, DIEA or NMM at a temperature ranging from about 20° C. toabout 150° C., preferably at room temperature, for a few hours, e.g. onehour to 24 h in a suitable solvent such as DMF, EtOH. The resultingester can then be hydrolysed to give compounds of Formula (IV) usingconditions and methods well known to those skilled in the art, such asbut not limited to the use of a metal hydroxide, e.g. lithium hydroxide,sodium hydroxide or potassium hydroxide, in a suitable solvent such asTHF, methanol or water or mixtures thereof, at a temperature rangingfrom about 20° C. to about 50° C., preferably at room temperature, for afew hours, e.g. one hour to 24 h.

Alternatively, compounds of Formula (IV), wherein R^(a) and R^(b) aredefined as above, can be obtained as outlined in Scheme 8 when R^(a) isa tertiary amino group. The first step consists in the reaction of acommercially available secondary amine (R^(a)—H) with a commerciallyavailable 4-fluorobenzonitrile derivative in the presence or absence ofbases such as TEA, DIEA or NMM at a temperature rising from about 20° C.to about 150° C., preferably at 60° C., for a few hours, preferably 8 h,neat or in a suitable solvent such as DMF, or EtOH. The resultingnitrile derivative can then be transformed into the corresponding esterderivative using conditions and methods well known to those skilled inthe art, such as but not limited to the use of a mineral acid such asHCl in an alcohol such as MeOH, EtOH, preferably MeOH at a temperaturerising from about 20° C. to about 100° C., preferably 60° C. for a fewhours, preferably 24 h. The resulting ester can then be hydrolysed togive compounds of Formula (IV) by the use of a metal hydroxide, e.g.lithium hydroxide, sodium hydroxide or potassium hydroxide, in asuitable solvent such as THF, methanol or water or mixtures thereof, ata temperature rising from 20° C. to 50° C., preferably at roomtemperature, for a few hours, e.g. one hour to 24 h.

The compounds of Formula (IV), wherein R^(b) is defined as above andwherein R^(a) is a tertiary amino group, can be obtained as outlined inScheme 9. The first step consists of the reaction of a commerciallyavailable secondary amine (R^(a)—H) with a 4-bromobenzoic esterderivative in the presence of a palladium source such as Pd(OAc)₂, aligand such as BINAP and a base such as Cs₂CO₃, K₂CO₃, tBuOK, tBuONa,TEA, DIEA or NMM. The reaction is preferrably performed at a temperaturerising from about 20° C. to about 150° C., preferably between 80° C. and120° C., for a few hours, e.g. one hour to 24 h in a suitable solventsuch as DMF, EtOH or dioxane. The resulting ester can then be hydrolysedto give compounds of Formula (IV) using conditions and methods wellknown to those skilled in the art, such as but not limited to the use ofa metal hydroxide, e.g. lithium hydroxide, sodium hydroxide or potassiumhydroxide, in a suitable solvent such as THF, methanol or water ormixtures thereof, at a temperature ranging from about 20° C. to about50° C., preferably at room temperature, for a few hours, e.g. one hourto 24 h.

The method for preparing the compounds of Formula (IV) selected below:

-   3-nitro-4-piperidin-1-ylbenzoic acid-   4-morpholin-4-yl-3-nitrobenzoic acid-   4-piperidin-1-ylbenzoic acid-   4-[cyclohexyl(methyl)amino]-3-nitrobenzoic acid-   4-(2-methylpiperidin-1-yl)-3-nitrobenzoic acid-   4-(3,3-difluoropiperidin-1-yl)-3-nitrobenzoic acid-   3-nitro-4-pyrrolidin-1-ylbenzoic acid-   4-azepan-1-yl-3-nitrobenzoic acid    -   4-(2-methylpiperidin-1-yl)-3-(trifluoromethyl)benzoic acid-   4-(2,5-dimethylpyrrolidin-1-yl)-3-nitrobenzoic acid-   4-[bis(2-methoxyethyl)amino]-3-nitrobenzoic acid-   3-cyano-4-(2-methylpiperidin-1-yl)benzoic acid-   6-(2-methylpiperidin-1-yl)nicotinic acid-   5-methyl-6-(2-methylpiperidin-1-yl)nicotinic acid-   6-[2-(methoxymethyl)pyrrolidin-1-yl]-5-methylnicotinic acid-   3-methyl-4-piperidin-1-ylbenzoic acid-   4-piperidin-1-yl-3-(trifluoromethyl)benzoic acid    is more particularly described in the Examples.

Alternatively, compounds of Formula (IV), wherein R^(b) is defined asabove and wherein R^(a) is an aryl or heteroaryl group, can be obtainedas outlined in Scheme 10. The first step consists of coupling acommercially available 4-bromobenzoic ester derivative with acommercially available aryl- or heteroaryl-boronic acid (R^(a)—B(OH)₂),a commercially available aryl- or heteroaryl-boronic pinacol ester or acommercially available aryl- or heteroaryl-tributylstannane, in thepresence of a source of palladium such astetrakis(triphenylphosphine)palladium(0) in a suitable solvent such as amixture of toluene and water at a temperature rising from about 20° C.to about 150° C., preferably 120° C. for a few hours, preferably 1 to 14hours in the presence or absence of a base such as TEA, DIEA, NaHCO₃ orK₂CO₃. The 4-arylbenzoic ester derivative obtained can then behydrolysed to give compounds of Formula (IV) by the use of a metalhydroxide, e.g. lithium hydroxide, sodium hydroxide or potassiumhydroxide, in a suitable solvent such as THF, methanol or water ormixtures thereof, at a temperature ranging from about 20° C. to about50° C., preferably at room temperature, for a few hours, e.g. one hourto 24 h.

Alternatively, compounds of Formula (IV), wherein R^(a) and R^(b) aredefined as above can be obtained as outlined in Scheme 11 when R^(a) isan aryl or heteroaryl group. The first step consists of theesterification of a commercially available 4-boronicester-benzoic acidderivative using conditions and methods well known to those skilled inthe art, such as thionyl chloride in methanol. This4-boronicester-benzoic ester derivative intermediate can then be reactedwith an halogenated- or trifluoromethanesulfonyl-aromatic orheteroaromatic in the presence of a source of palladium such asPd(PPh₃)₄ in a suitable solvent such as a mixture of toluene and waterat a temperature rising from about 20° C. to about 150° C., preferably120° C. for a few hours, preferably 1 to 14 hours in the presence orabsence of a base such as TEA, DIEA, NaHCO₃ or K₂CO₃. The 4-arylbenzoicester derivative obtained can then be hydrolysed to give compounds ofFormula (IV) by the use of a metal hydroxide, e.g. lithium hydroxide,sodium hydroxide or potassium hydroxide, in a suitable solvent such asTHF, methanol or water or mixtures thereof, at a temperature rangingfrom about 20° C. to about 50° C., preferably at room temperature, for afew hours, e.g. one hour to 24 h.

Alternatively, compounds of Formula (V), wherein R^(a) is an aryl orheteroaryl group, can be obtained as outlined in Scheme 12. The firststep consists in bromination of 4-bromo-3-methyl-benzoic esterderivatives using conditions known by one skill in the art such as NBSin the presence of AIBN, in a suitable solvent such as CHCl₃ or DCE at atemperature rising from room temperature to about 120°, preferablybetween 60° C. and 100° C. for a few hours. The benzylic bromine of thisintermediate can then be substituted by an alkylcarboxylate such as anacetate by reaction with AcONa in AcOH, at a temperature rising fromroom temperature to about 150°, preferably between 80° C. and 120° C.for a few hours, preferably between 5 and 24 hours. This intermediatecan then be coupled with a commercially available aryl- orheteroaryl-boronic acid (R^(a)—B(OH)₂), a commercially available aryl-or heteroaryl-boronic pinacol ester or a commercially available aryl- orheteroaryl-tributylstannane, in the presence of a source of palladiumsuch as tetrakis(triphenylphosphine)palladium(0) in a suitable solventsuch as a mixture of toluene and water at a temperature rising fromabout 20° C. to about 150° C., preferably 120° C. for a few hours,preferably 1 to 14 hours in the presence or absence of a base such asTEA, DIEA, NaHCO₃ or K₂CO₃. The 4-arylbenzoic ester derivative obtainedcan then be hydrolysed to give compounds of Formula (V) by the use of ametal hydroxide, e.g. lithium hydroxide, sodium hydroxide or potassiumhydroxide, in a suitable solvent such as THF, methanol or water ormixtures thereof, at a temperature rising from about 20° C. to about 50°C., preferably at room temperature, for a few hours, e.g. one hour to 24h.

The method for preparing the compounds of Formula (V) selected below:

-   2,2′-dimethyl-1,1′-biphenyl-4-carboxylic acid-   2-methyl-2′-(trifluoromethyl)biphenyl-4-carboxylic acid-   3-methyl-4-(4-methyl-3-thienyl)benzoic acid-   2′-methyl-2-nitro-1,1′-biphenyl-4-carboxylic acid-   2′-methoxy-2-methylbiphenyl-4-carboxylic acid-   2-methoxy-2′-methylbiphenyl-4-carboxylic acid-   2′,4′-dimethoxy-2-methylbiphenyl-4-carboxylic acid-   3-methoxy-4-(4-methyl-3-thienyl)benzoic acid-   4-(3,5-dimethylisoxazol-4-yl)-3-methylbenzoic acid-   3-methyl-4-(2-methylpyridin-3-yl)benzoic acid    -   2-(hydroxymethyl)-2′-methylbiphenyl-4-carboxylic acid        is more particularly described in the Examples.

If the above set out general synthetic methods are not applicable forthe obtention of compounds of Formula (I), suitable methods ofpreparation known by a person skilled in the art should be used.

The pharmaceutically acceptable cationic salts of compounds of thepresent invention are readily prepared by reacting the acid forms withan appropriate base, usually one equivalent, in a co-solvent. Typicalbases are sodium hydroxide, sodium methoxide, sodium ethoxide, sodiumhydride, potassium hydroxide, potassium methoxide, magnesium hydroxide,calcium hydroxide, benzathine, choline, diethanolamine, ethylenediamine,meglumine, benethamine, diethylamine, piperazine and tromethamine. Thesalt is isolated by concentration to dryness or by addition of anon-solvent. In some cases, salts can be prepared by mixing a solutionof the acid with a solution of the cation (sodium ethylhexanoate,magnesium oleate), employing a solvent in which the desired cationicsalt precipitates, or can be otherwise isolated by concentration andaddition of a non-solvent.

According to a further general process, compounds of Formula (I) can beconverted to alternative compounds of Formula (I), employing suitableinterconversion techniques well known by a person skilled in the art.

In general, the synthesis pathways for any individual compound ofFormula (I) will depend on the specific substitutents of each moleculeand upon the ready availability of intermediates necessary; again suchfactors being appreciated by those of ordinary skill in the art. For allthe protection and deprotection methods, see Philip J. Kocienski, in“Protecting Groups”, Georg Thieme Verlag Stuttgart, New York, 1994 and,Theodora W. Greene and Peter G. M. Wuts in “Protective Groups in OrganicSynthesis”, Wiley Interscience, 3^(rd) Edition 1999.

Compounds of this invention can be isolated in association with solventmolecules by crystallization from evaporation of an appropriate solvent.The pharmaceutically acceptable acid addition salts of the compounds ofFormula (I), which contain a basic center, may be prepared in aconventional manner. For example, a solution of the free base may betreated with a suitable acid, either neat or in a suitable solution, andthe resulting salt isolated either by filtration or by evaporation undervacuum of the reaction solvent. Pharmaceutically acceptable baseaddition salts may be obtained in an analogous manner by treating asolution of compound of Formula (I) and, which contain an acid center,with a suitable base. Both types of salts may be formed orinterconverted using ion-exchange resin techniques.

Depending on the conditions used, the reaction times are generallybetween a few minutes and 14 days, and the reaction temperature isbetween about −30° C. and 140° C., normally between −10° C. and 90° C.,in particular between about 0° C. and about 70° C.

Compounds of the formula (I) can furthermore be obtained by liberatingcompounds of the formula (I) from one of their functional derivatives bytreatment with a solvolysing or hydrogenolysing agent.

Preferred starting materials for the solvolysis or hydrogenolysis arethose which conform to the formula (I), but contain correspondingprotected amino and/or hydroxyl groups instead of one or more free aminoand/or hydroxyl groups, preferably those which carry an amino-protectinggroup instead of an H atom bonded to an N atom, in particular thosewhich carry an R′—N group, in which R′ denotes an amino-protectinggroup, instead of an HN group, and/or those which carry ahydroxyl-protecting group instead of the H atom of a hydroxyl group, forexample those which conform to the formula (I), but carry a —COOR″group, in which R″ denotes a protecting group, instead of a —COOH group.

It is also possible for a plurality of—identical or different—protectedamino and/or hydroxyl groups to be present in the molecule of thestarting material. If the protecting groups present are different fromone another, they can in many cases be cleaved off selectively.

The term “amino-protecting group” is known in general terms and relatesto groups which are suitable for protecting (blocking) an amino groupagainst chemical reactions, but which are easy to remove after thedesired chemical reaction has been carried out elsewhere in themolecule. Typical of such groups are, in particular, unsubstituted orsubstituted acyl, aryl, aralkoxymethyl or aralkyl groups. Since theamino-protecting groups are removed after the desired reaction (orreaction sequence), their type and size are furthermore not crucial;however, preference is given to those having 1-20, in particular 1-8,carbon atoms. The term “acyl group” is to be understood in the broadestsense in connection with the present process. It includes acyl groupsderived from aliphatic, araliphatic, aromatic or hetero-cycliccarboxylic acids or sulfonic acids, and, in particular, alkoxy-carbonyl,aryloxycarbonyl and especially aralkoxycarbonyl groups. Examples of suchacyl groups are alkanoyl, such as acetyl, propionyl and butyryl;aralkanoyl, such as phenylacetyl; aroyl, such as benzoyl and tolyl;aryloxyalkanoyl, such as POA; alkoxycarbonyl, such as methoxy-carbonyl,ethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, BOC(tert-butoxy-carbonyl) and 2-iodoethoxycarbonyl; aralkoxycarbonyl, suchas CBZ (“carbo-benz-oxy”), 4-methoxybenzyloxycarbonyl and FMOC; andaryl-sulfonyl, such as Mtr. Preferred amino-protecting groups are BOCand Mtr, furthermore CBZ, Fmoc, benzyl and acetyl.

The term “hydroxyl-protecting group” is likewise known in general termsand relates to groups which are suitable for protecting a hydroxyl groupagainst chemical reactions, but are easy to remove after the desiredchemical reaction has been carried out elsewhere in the molecule.Typical of such groups are the above-mentioned unsubstituted orsubstituted aryl, aralkyl or acyl groups, furthermore also alkyl groups.The nature and size of the hydroxyl-protecting groups are not crucialsince they are removed again after the desired chemical reaction orreaction sequence; preference is given to groups having 1-20, inparticular 1-10, carbon atoms.

Examples of hydroxyl-protecting groups are, inter alia, benzyl,4-methoxybenzyl, p-nitro-benzoyl, p-toluenesulfonyl, tert-butyl andacetyl, where benzyl and tert-butyl are particularly preferred.

The compounds of the formula (I) are liberated from their functionalderivatives—depending on the protecting group used—for example usingstrong acids, advantageously using TFA or perchloric acid, but alsousing other strong inorganic acids, such as hydrochloric acid orsulfuric acid, strong organic carboxylic acids, such as trichloroaceticacid, or sulfonic acids, such as benzene- or p-toluenesulfonic acid. Thepresence of an additional inert solvent is possible, but is not alwaysnecessary. Suitable inert solvents are preferably organic, for examplecarboxylic acids, such as acetic acid, ethers, such as tetrahydrofuranor dioxane, amides, such as DMF, halogenated hydrocarbons, such asdichloromethane, furthermore also alcohols, such as methanol, ethanol orisopropanol, and water. Mixtures of the above-mentioned solvents arefurthermore suitable. TFA is preferably used in excess without additionof a further solvent, and perchloric acid is preferably used in the formof a mixture of acetic acid and 70% perchloric acid in the ratio 9:1.The reaction temperatures for the cleavage are advantageously betweenabout 0 and about 50° C., preferably between 15 and 30° C. (roomtemperature).

The BOC, OtBu and Mtr groups can, for example, preferably be cleaved offusing TFA in dichloromethane or using approximately 3 to 5N HCl indioxane at 15-30° C., and the FMOC group can be cleaved off using anapproximately 5 to 50% solution of dimethylamine, diethylamine orpiperidine in DMF at 15-30° C.

Protecting groups which can be removed hydrogenolytically (for exampleCBZ, benzyl or the liberation of the amidino group from the oxadiazolederivative thereof) can be cleaved off, for example, by treatment withhydrogen in the presence of a catalyst (for example a noble-metalcatalyst, such as palladium, advantageously on a support, such ascarbon). Suitable solvents here are those indicated above, inparticular, for example, alcohols, such as methanol or ethanol, oramides, such as DMF. The hydrogenolysis is generally carried out attemperatures between about 0 and 100° C. and pressures between about 1and 200 bar, preferably at 20-30° C. and 1-10 bar. Hydrogenolysis of theCBZ group succeeds well, for example, on 5 to 10% Pd/C in methanol orusing ammonium formate (instead of hydrogen) on Pd/C in methanol/DMF at20-30° C.

Examples of suitable inert solvents are hydrocarbons, such as hexane,petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons,such as trichloroethylene, 1,2-dichloroethane, tetrachloromethane,tri-fluoro-methylbenzene, chloroform or dichloromethane; alcohols, suchas methanol, ethanol, isopropanol, n-propanol, n-butanol ortert-butanol; ethers, such as diethyl ether, diisopropyl ether,tetrahydrofuran (THF) or dioxane; glycol ethers, such as ethylene glycolmonomethyl or monoethyl ether or ethylene glycol dimethyl ether(diglyme); ketones, such as acetone or butanone; amides, such asacetamide, dimethylacetamide, N-methylpyrrolidone (NMP) ordimethyl-formamide (DMF); nitriles, such as acetonitrile; sulfoxides,such as dimethyl sulfoxide (DMSO); carbon disulfide; carboxylic acids,such as formic acid or acetic acid; nitro compounds, such asnitromethane or nitrobenzene; esters, such as ethyl acetate, or mixturesof the said solvents.

Esters can be saponified, for example, using acetic acid or using LiOH,NaOH or KOH in water, water/THF, water/THF/ethanol or water/dioxane, attemperatures between 0 and 100° C.

Free amino groups can furthermore be acylated in a conventional mannerusing an acid chloride or anhydride or alkylated using an unsubstitutedor substituted alkyl halide or reacted with CH₃—C(═NH)—OEt,advantageously in an inert solvent, such as dichloromethane or THFand/or in the presence of a base, such as triethylamine or pyridine, attemperatures between −60° C. and +30° C.

Therefore, the invention also relates to the preparation of thecompounds of formula (I), and salts thereof, characterized in that

a) a Compound of Formula A

wherein W, R^(a) and R^(b) have the meanings given above, and T is OH,or a leaving group, such as Cl, Br, I, imidazolyl, pentafluorophenoxy orthe product of the reaction of isobutyl chloroformate with formula A,wherein T is OH, is reacted witha compound of formula B

wherein R¹, R² and R^(c) have the meanings given above, preferably inthe presence of a solvent and of a suitable base, such as an amine likeTEA, DIEA or NMM, or in case T is OH, in the presence of a suitablecondensation reagent, such as EDC and the resulting product is cyclized,preferably in the presence of an amine, such as DIEA, TEA ortetrabutylamonium fluorideand optionally a base or acid of the formula (I) is converted into oneof its salts.

Throughout the specification, the term leaving group preferably denotesCl, Br, I or a reactively modified OH group, such as, for example, anactivated ester, an imidazolide or alkylsulfonyloxy having 1-6 carbonatoms (preferably methylsulfonyloxy or trifluoromethylsulfonyloxy) orarylsulfonyloxy having 6-10 carbon atoms (preferably phenyl- orp-tolylsulfonyloxy). Radicals of this type for activation of thecarboxyl group in typical acylation reactions are described in theliterature (for example in the standard works, such as Houben-Weyl,Methoden der organischen Chemie [Methods of Organic Chemistry],Georg-Thieme-Verlag, Stuttgart). Activated esters are advantageouslyformed in situ, for example through addition of HOBt orN-hydroxysuccinimide.

The formula (I) also encompasses the optically active forms(stereoisomers), the enantiomers, the racemates, the diastereomers andthe hydrates and solvates of these compounds. The term “solvates of thecompounds” is taken to mean adductions of inert solvent molecules ontothe compounds which form owing to their mutual attractive force.Solvates are, for example, mono- or dihydrates or alcoholates.

The term “pharmaceutically usable derivatives” is taken to mean, forexample, the salts of the compounds of the formula (I) and so-calledprodrug compounds.

The term “prodrug derivatives” is taken to mean compounds of the formula(I) which have been modified with, for example, alkyl or acyl groups,sugars or oligopeptides and which are rapidly cleaved in the organism toform the active compounds.

These also include biodegradable polymer derivatives of the compoundsaccording to the invention, as described, for example, in Int. J. Pharm.115, 61-67 (1995).

The Formula (I) also encompasses compounds wherein R^(a) is OA wherein Ais as above described.

The formula (I) also encompasses compounds wherein R^(c) is H.

The Formula (I) also encompasses compounds wherein Het denotes asaturated, unsaturated or aromatic heterocyclic ring having 1 to 4 Natoms.

The formula (I) also encompasses mixtures of the compounds of theformula (I), for example mixtures of two diastereomers, for example inthe ratio 1:1, 1:2, 1:3, 1:4, 1:5, 1:10, 1:100 or 1:1000.

These are particularly preferably mixtures of stereoisomeric compounds.

In a preferred embodiment, the invention provides compounds of Formula(I′)

Wherein R^(a), R^(b), are as above defined

R¹ is H or F R^(c) is CF₃ or Me

In another preferred embodiment, the invention provides compounds ofFormula (I″)

Wherein R^(b), R^(c) and R¹ are as defined under Formula (I) and whereinR^(x) denotes Hal, A, CH₂OA, CH₂N(R₃)₂, OR³, N(R³)₂, NO₂, N(SO₂)Me)₂,CN, COOR³, CF₃, OCF₃, CON(R³)₂, NR³COA, NR³CON(R³)₂, NR³SO₂A, COR³,SO₂N(R³)₂, SOA or SO₂A, phenyl, pyridyl, —[C(R³)₂]_(n)—COOR³ or—C[C(R³)₂], —CON(R³)₂. Preferrably R^(x) is in such position that itlimits the rotation of the ring Z with respect to the ring bearingR^(b), by the meaning of steric hindrance or electrostatic interactionswith R^(b). R^(x) is preferrably an alkyl or an alkoxy chain containing1 to 5 carbon atoms. R^(x) is most preferrably attached to the atomadjacent to the atom which links the ring Z to the rest of the molecule.R^(x) is most preferrably —CH₃, —C₂H₅, F, Cl, —OCH₃, —CH₂OCH₃ and R^(b)is simultaneously —CH₃, —C₂H₅, F, Cl, —OCH₃, —OC₂H₅, —CH₂OCH₃—CH₂OH,—CH₂N(CH₃)₂, CF₃.

Preference is given to the compounds of the present invention selectedfrom the following group I1 to I79:

and pharmaceutically usable derivatives, solvates, salts andstereoisomers thereof, including mixtures thereof in all ratios.

For all radicals which occur more than once, their meanings areindependent of one another. Above and below, the radicals or parametersR^(a), R^(b), R^(c), R¹, R², R³, W, T, X, A, Ar, Het and n have themeaning indicated under the formula I, unless expressly statedotherwise.

A denotes alkyl, is unbranched (linear) or branched, and has 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. A preferably denotes methyl,furthermore ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl ortert-butyl, furthermore also pentyl, 1-, 2- or 3-methylbutyl, 1,1-, 1,2-or 2,2-dimethyl-propyl, 1-ethylpropyl, hexyl, 1-, 2-, 3- or4-methylpentyl, 1,1-, 1,2-, 1,3-, 2,2-, 2,3- or 3,3-dimethylbutyl, 1- or2-ethylbutyl, 1-ethyl-1-methylpropyl, 1-ethyl-2-methylpropyl, 1,1,2- or1,2,2-trimethylpropyl, furthermore preferably, for example,trifluoromethyl. A very particularly preferably denotes alkyl having 1,2, 3, 4, 5 or 6 carbon atoms, preferably methyl, ethyl, propyl,isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl,trifluoromethyl, pentafluoroethyl, 1,1,1-trifluoroethyl. In a preferredembodiment A is perfluorated. A furthermore denotes—(CH₂)_(n)—O—(CH₂)_(n)OR³, —(CH₂)_(n)NR³(CH₂)₂N(R³)₂, especially—(CH₂)₂—O—(CH₂)₂OR³ or —(CH₂)₂NH(CH₂)₂N(R³)₂.

Cycloalkyl are cyclic alkyl containing 3 to 12 carbon atomes.

Cycloalkyl preferably denotes cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl or cycloheptyl.

Cycloalkylalkylene is a cycloalkyl group bond to the rest of themolecule via a carbon chain and having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10,11, 12, 13, 14, 15, 16, 17, or 18 carbon atoms.

Cycloalkylalkylene preferably denotes cyclopropylmethylene,cyclobutylmethylene, cyclopentylmethylene, cyclohexylmethylene orcycloheptylmethylene.

Alkylene is a bivalent carbon chain having 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, or 12 carbon atoms. Alkylene is preferably methylene, ethylene,propylene, butylene, pentylene or hexylene, furthermore branchedalkylene.

R^(a) is preferably Ar, Het or OA especially Ar or Het.

If Het denotes a N-Atom bearing saturated heterocycle, Het is preferablylinked to the rest of the molecule via an N-Atom. The alpha position isnext to this N-Atom.

R^(b) is preferably H, A, OH, OA, especially —OCH₃, —OCF₃, —CH₃, —NO₂,Hal, —CH₂OR³, —CH₂NHSO₂A, —NHSO₂A, —NH₂, —CH₂NHCOCH₃, —CH₂N(CH₃)₂,—CH₂NH₂, —NHCONH₂, —(CH₂)_(n)SO₂R³—N(SO₂A)₂, —CO₂R³, or —CF₃

The invention also encompasses compounds of Formula (I) wherein R^(b),OR³, CN, or Hal

R^(c) preferably denotes alkyl, polyfluoralkyl or perfluoralkyl,especially —CH₃, —C₂H₅ or —CF₃.

Compounds of formula (I), wherein R^(c) denotes H are preferred asintermediates for the synthesis of other compounds of formula I.

R³ is preferably A.

Hal is preferably F, Cl or Br and especially F or Cl.

Preferably, at least one of R¹ and R² denotes F or Cl.

-   -   R¹ preferably denotes F or H.    -   R¹ is preferably in para position to the group OR^(c).    -   R² is preferably F or H, especially H.

W preferably denotes CH.

-   -   n is preferably 0, 1, 2, 3, 4 or 5 and more preferably 0, 1, 2,        3 or 4.

An aromatic carbocyclic ring preferably denotes phenyl, naphthyl orbiphenyl.

Ar denotes, for example, phenyl, o-, m- or p-tolyl, o-, m- orp-ethylphenyl, o-, m- or p-propyl-phenyl, o-, m- or p-isopropylphenyl,o-, m- or p-tert-butylphenyl, o-, m- or p-hydroxyphenyl, o-, m- orp-nitrophenyl, o-, m- or p-aminophenyl, o-, m- orp-(N-methylamino)phenyl, o-, m- or p-(N-methylaminocarbonyl)phenyl, o-,m- or p-acetamidophenyl, o-, m- or p-methoxyphenyl, o-, m- orp-ethoxyphenyl, o-, m- or p-ethoxycarbonylphenyl, o-, m- orp-(N,N-dimethylamino)phenyl, o-, m- orp-(N,N-dimethylaminocarbonyl)phenyl, o-, m- or p-(N-ethylamino)phenyl,o-, m- or p-(N,N-diethylamino)phenyl, o-, m- or p-fluorophenyl, o-, m-or p-bromophenyl, o-, m- or p-chloro-phenyl, o-, m- orp-(methylsulfonamido)phenyl, o-, m- or p-(methylsulfonyl)phenyl, o, m orp-amino-sulfanyl-phenyl, o-, m- or p-phenoxyphenyl, further preferably2,3-, 2,4-, 2,5-, 2,6-, 3,4- or 3,5-dimethylphenyl, 2,3-, 2,4-, 2,5-,2,6-, 3,4- or 3,5-difluorophenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- or3,5-dichlorophenyl, 2,3-, 2,4-, 2,5-, 2,6-, 3,4- or 3,5-dibromophenyl,2,4- or 2,5-dinitrophenyl, 2,5- or 3,4-dimethoxyphenyl,3-nitro-4-chlorophenyl, 3-amino-4-chloro-, 2-amino-3-chloro-,2-amino-4-chloro-, 2-amino-5-chloro- or 2-amino-6-chlorophenyl,2-nitro-4-N,N-dimethylamino- or 3-nitro-4-N,N-dimethylaminophenyl,2,3-diaminophenyl, 2,3,4-, 2,3,5-, 2,3,6-, 2,4,6- or3,4,5-trichlorophenyl, 2,4,6-trimethoxyphenyl,2-hydroxy-3,5-dichlorophenyl, p-iodophenyl, 3,6-dichloro-4-aminophenyl,4-fluoro-3-chlorophenyl, 2-fluoro-4-bromophenyl,2,5-difluoro-4-bromo-phenyl, 3-bromo-6-methoxyphenyl,3-chloro-6-methoxyphenyl, 3-chloro-4-acetamidophenyl,3-fluoro-4-methoxyphenyl, 3-amino-6-methylphenyl,3-chloro-4-acetamidophenyl or 2,5-dimethyl-4-chlorophenyl.

Ar preferably denotes, for example, phenyl which is unsubstituted ormonosubstituted, disubstituted or trisubstituted by A, Hal, OR³, CF₃,OCF₃, NO₂ and/or CN. If Ar is phenyl, it is preferably substituted in 2′position, i.e. in ortho-position to the oxadiazole bearing moiety. Ar ispreferably substituted by A, OR³, CF₃ OCF₃.

Ar particularly preferably denotes, for example, phenyl which isunsubstituted or monosubstituted or disubstituted preferablymonosubstituted, by F, OCH₃, CH₃, CF₃, phenyl and/or pyridyl, such as,for example, 2′-methoxy-phenyl-, 2′-trifluoromethyl-phenyl-(aryl bearingat least a 2′ substituent), 2′-chloro-phenyl, 2′,6′-dimethyl-phenyl- or2′-alkyl-phenyl-, preferably 2′-methyl-phenyl.

Ar very particularly preferably denotes one of the following groups:

preferably

Wherein X′ and X² are independently of one another F, Cl, —OCH₃, —CH₃,—C₂H₅, —CF₃, —OCF₃, —O-isoPropyl, —O-isobutyl, —OCH₂CN,—OCH₂cyclopropyl, —CH₂OH, —CH₂O-isoPropyl, —CH₂O-isobutyl,—CH₂OCH₂cyclopropyl, —CH₂NMe₂, —CH₂OC₂H₅, —NHCOMe, —NHCOEt, —NHSO₂NMe₂,—NHSO₂propyl, —CH₂-morpholine, —CH₂pirolidine, —CH₂NHMe, —SO₂Me,—CH₂SO₂Me, —C≡C—CH₂OMe, —(CH₂)₃OMe, —O(CH₂)₂OMe, —CO₂H, —OH, —NO₂, —CN,—NHSO₂CH₃, and/or phenyl or pyridyl or piperidine or morpholine, whichis preferably unsubstituted,

More preferably, X¹ and X² denote independently of one another —F,—OCH₃, —CH₃, —CF₃, —OCF₃, —OH, —NO₂, —CN, and/or phenyl or pyridyl,which is preferably unsubstituted.

Het is preferably a 6 to 14 membered ring system and denotes, notwithstanding further substitutions, for example, 2- or 3-furyl, 2- or3-thienyl, 1-, 2- or 3-pyrrolyl, 1-, 2-, 4- or 5-imidazolyl, 1-, 3-, 4-or 5-pyrazolyl, 2-, 4- or 5-oxazolyl, 3-, 4- or 5-isoxazolyl, 2-, 4- or5-thiazolyl, 3-, 4- or 5-isothiazolyl, 2-, 3- or 4-pyridyl, 2-, 4-, 5-or 6-pyrimidinyl, furthermore preferably 1,2,3-triazol-1-, -4- or -5-yl,1,2,4-triazol-1-, -3- or -5-yl, 1- or 5-tetrazolyl, 1,2,3-oxadiazol-4-or -5-yl, 1,2,4-oxadiazol-3- or -5-yl, 1,3,4-thiadiazol-2- or -5-yl,1,2,4-thiadiazol-3- or -5-yl, 1,2,3-thiadiazol-4- or -5-yl, 3- or4-pyridazinyl, pyrazinyl, 1-, 2-, 3-, 4-, 5-, 6- or 7-indolyl,indazolyl, 4- or 5-isoindolyl, 1-, 2-, 4- or 5-benzimidazolyl, 1-, 3-,4-, 5-, 6- or 7-benzopyrazolyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 3-,4-, 5-, 6- or 7-benzisoxazolyl, 2-, 4-, 5-, 6- or 7-benzo-thiazolyl, 2-,4-, 5-, 6- or 7-benzisothiazolyl, 4-, 5-, 6- or7-benz-2,1,3-oxadiazolyl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-,4-, 5-, 6-, 7- or 8-isoquinolyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-,4-, 5-, 6-, 7- or 8-quinazolinyl, 5- or 6-quinoxalinyl, 2-, 3-, 5-, 6-,7- or 8-2H-benzo-1,4-oxazinyl, furthermore preferably1,3-benzodioxol-5-yl, 1,4-benzodioxane-6-yl, 2,1,3-benzothiadiazol-4- or-5-yl or 2,1,3-benzoxadiazol-5-yl.

The heterocyclic radicals may also be partially or fully hydrogenated.

Het can thus also denote, for example, 2,3-dihydro-2-, -3-, -4- or-5-furyl, 2,5-dihydro-2-, -3-, -4- or -5-furyl, tetrahydro-2- or-3-furyl, 1,3-dioxolan-4-yl, tetrahydro-2- or -3-thienyl,2,3-dihydro-1-, -2-, -3-, -4- or -5-pyrrolyl, 2,5-dihydro-1-, -2-, -3-,-4- or -5-pyrrolyl, 1-, 2- or 3-pyrrolidinyl, tetrahydro-1-, -2- or-4-imidazolyl, 2,3-dihydro-1-, -2-, -3-, -4- or -5-pyrazolyl,tetrahydro-1-, -3- or -4-pyrazolyl, 1,4-dihydro-1-, -2-, -3- or-4-pyridyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5- or -6-pyridyl, 1-,2-, 3- or 4-piperidinyl, 2-, 3- or 4-morpholinyl, tetrahydro-2-, -3- or-4-pyranyl, 1,4-dioxaneyl, 1,3-dioxane-2-, -4- or -5-yl, hexahydro-1-,-3- or -4-pyridazinyl, hexahydro-1-, -2-, -4- or -5-pyrimidinyl, 1-, 2-or 3-piperazinyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7- or-8-quinolyl, 1,2,3,4-tetrahydro-1-, -2-, -3-, -4-, -5-, -6-, -7- or-8-isoquinolyl, 2-, 3-, 5-, 6-, 7- or8-3,4-dihydro-2H-benzo-1,4-oxazinyl, furthermore preferably2,3-methylenedioxyphenyl, 3,4-methylenedioxyphenyl,2,3-ethylenedioxyphenyl, 3,4-ethylenedioxyphenyl,3,4-(difluoro-methylenedioxy)phenyl, 2,3-dihydrobenzofuran-5- or -6-yl,2,3-(2-oxomethylenedioxy)phenyl or also3,4-dihydro-2H-1,5-benzodioxepin-6- or -7-yl, furthermore preferably2,3-dihydrobenzo-furanyl or 2,3-dihydro-2-oxofuranyl.

Het very particularly denotes one of the following groups:

wherein X¹, X², and R³ are as defined above.

The compounds of the formula (I) can have one or more centres ofchirality and can therefore occur in various stereoisomeric forms. Theformula (I) covers all these forms.

Accordingly, the invention relates, in particular, to compounds ofFormula (I) and its use, in which at least one of the said radicals hasone of the preferred meanings indicated above. Some preferred groups ofcompounds can be expressed by the following sub-formulae Ia to Ih, whichconform to the formula I and in which the radicals not designated ingreater detail have the meaning indicated under the formula (I), but inwhich

-   -   in Ia R^(a) Ar or Het.    -   in Ib R^(a) is heterocycloalkyl or heteroaryl which both may be        unsubstituted or monosubstituted or disubstituted, preferably        monosubstituted, by F, OCH₃, CH₃, CF₃, such as, for example,        morpholino-4-yl or 2-methyl-piperidin-1-yl,    -   in Ic        -   R¹ and R² denote H.    -   in Id R^(a) is heterocycloalkyl such as morpholino-4-yl and        2-methyl-piperidin-1-yl        -   R^(b) is polyfluoroalkyl,        -   R^(c) is selected from alkyl, polyfluoroalkyl,    -   in Ie R^(a) is heterocycloalkyl such as morpholino-4-yl        -   R^(b) is amino,        -   R^(c) is selected from alkyl,    -   in If R^(a) is heterocycloalkyl such as morpholino-4-yl,        2-methyl-piperidin-1-yl, 3,3-difluoropiperidin-1-yl and        piperidin-1-yl        -   R^(b) is nitro,        -   R^(c) is alkyl,    -   in Ig R^(a) is heterocycloalkyl such as morpholino-4-yl,        2-methyl-piperidin-1-yl, 3,3-difluoropiperidin-1-yl and        piperidin-1-yl        -   R^(b) is nitro,        -   R^(c) is polyfluoroalkyl,    -   in Ih R^(a) is pyrrolidin-1-yl        -   R^(b) is nitro,        -   R^(c) is selected from alkyl, polyfluoroalkyl,    -   in Ii R^(a) is aryl or heteroaryl such as tol-2-yl,        2-trifluoromethylphen-1-yl and 4-methylthien-3-yl        -   R^(b) is selected from alkyl, nitro,        -   R^(c) is selected from alkyl, polyfluoroalkyl,            and pharmaceutically usable derivatives, solvates, salts and            stereoisomers thereof, including mixtures thereof in all            ratios.

The compounds of the formula (I) and also the starting materials for thepreparation thereof are, in addition, prepared by methods known per se,as described in the literature (for example in the standard works, suchas Houben-Weyl, Methoden der organischen Chemie [Methods of OrganicChemistry], Georg-Thieme-Verlag, Stuttgart), under reaction conditionswhich are known and suitable for the said reactions. For all theprotection and deprotection methods, see Philip J. Kocienski, in“Protecting Groups”, Georg Thieme Verlag Stuttgart, New York, 1994 and,Theodora W. Greene and Peter G. M. Wuts in “Protective Groups in OrganicSynthesis”, Wiley Interscience, 3^(rd) Edition 1999.

Use can also be made here of variants which are known per se, but arenot mentioned here in greater detail.

If desired, the starting materials can also be formed in situ so thatthey are not isolated from the reaction mixture, but instead areimmediately converted further into the compounds of the formula (I).

The starting compounds for the preparation of compounds of formula (I)are generally known. If they are novel, they can, however, be preparedby methods known per se.

The reactions are preferably carried out in an inert solvent.

Examples of suitable inert solvents are hydrocarbons, such as hexane,petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons,such as trichloroethylene, 1,2-dichloroethane, tetrachloromethane,chloroform or dichloromethane; alcohols, such as methanol, ethanol,isopropanol, n-propanol, n-butanol or tert-butanol; ethers, such asdiethyl ether, diisopropyl ether, tetrahydrofuran (THF) or dioxane;glycol ethers, such as ethylene glycol monomethyl or monoethyl ether orethylene glycol dimethyl ether (diglyme); ketones, such as acetone orbutanone; amides, such as acetamide, dimethylacetamide ordimethylformamide (DMF); nitriles, such as acetonitrile; sulfoxides,such as dimethyl sulfoxide (DMSO); carbon di-sulfide; carboxylic acids,such as formic acid or acetic acid; nitro compounds, such asnitromethane or nitrobenzene; esters, such as ethyl acetate, or mixturesof the said solvents.

Pharmaceutical Salts and Other Forms

The said compounds of the formula (I) can be used in their finalnon-salt form. On the other hand, the present invention also relates tothe use of these compounds in the form of their pharmaceuticallyacceptable salts, which can be derived from various organic andinorganic acids and bases by procedures known in the art.Pharmaceutically acceptable salt forms of the compounds of the formula(I) are for the most part prepared by conventional methods. If thecompound of the formula (I) contains an acidic center, such as acarboxyl group, one of its suitable salts can be formed by reacting thecompound with a suitable base to give the corresponding base-additionsalt. Such bases are, for example, alkali metal hydroxides, includingpotassium hydroxide, sodium hydroxide and lithium hydroxide; alkalineearth metal hydroxides, such as barium hydroxide and calcium hydroxide;alkali metal alkoxides, for example sodium- or potassiumethoxide andsodium or potassiumpropoxide, alkalihydrides, such as sodium- orpotassiumhydride; and various organic bases, such as piperidine,diethanolamine and N-methyl-glutamine, benzathine, choline,diethanolamine, ethylenediamine, meglumine, benethamine, diethylamine,piperazine and tromethamine. The aluminium salts of the compounds of theformula (I) are likewise included. In the case of certain compounds ofthe formula (I), which contain a basic center, acid-addition salts canbe formed by treating these compounds with pharmaceutically acceptableorganic and inorganic acids, for example hydrogen halides, such ashydrogen chloride, hydrogen bromide or hydrogen iodide, other mineralacids and corresponding salts thereof, such as sulfate, nitrate orphosphate and the like, and alkyl- and monoaryl-sulfonates, such asethanesulfonate, toluenesulfonate and benzene-sulfonate, and otherorganic acids and corresponding salts thereof, such as acetate,trifluoro-acetate, tartrate, maleate, succinate, citrate, benzoate,salicylate, ascorbate and the like. Accordingly, pharmaceuticallyacceptable acid-addition salts of the compounds of the formula (I)include the following: acetate, adipate, alginate, arginate, aspartate,benzoate, benzene-sulfonate (besylate), bisulfate, bisulfite, bromide,butyrate, camphorate, camphor-sulfonate, caprylate, chloride,chlorobenzoate, citrate, cyclo-pentane-propionate, digluconate,dihydrogen-phosphate, dinitrobenzoate, dodecyl-sulfate, ethanesulfonate,fumarate, galacterate (from mucic acid), galacturonate, glucoheptanoate,gluconate, glutamate, glycerophosphate, hemisuccinate, hemisulfate,heptanoate, hexanoate, hippurate, hydro-chloride, hydrobromide,hydroiodide, 2-hydroxy-ethane-sulfonate, iodide, isethionate,isobutyrate, lactate, lactobionate, malate, maleate, malonate,mandelate, metaphosphate, methanesulfonate, methylbenzoate,mono-hydrogen-phosphate, 2-naphthalenesulfonate, nicotinate, nitrate,oxalate, oleate, palmoate, pectinate, persulfate, phenylacetate,3-phenylpropionate, phosphate, phosphonate, phthalate, but this does notrepresent a restriction. Both types of salts may be formed orinterconverted preferably using ion-exchange resin techniques.

Furthermore, the base salts of the compounds of the formula (I) includealuminium, ammonium, calcium, copper, iron(III), iron(II), lithium,magnesium, manganese(III), manganese(II), potassium, sodium and zincsalts, but this is not intended to represent a restriction. Of theabove-mentioned salts, preference is given to ammonium; the alkali metalsalts sodium and potassium, and the alkaline earth metal salts calciumand magnesium. Salts of the compounds of the formula (I) which arederived from pharmaceutically acceptable organic non-toxic bases includesalts of primary, secondary and tertiary amines, substituted amines,also including naturally occurring substituted amines, cyclic amines,and basic ion exchanger resins, for example arginine, betaine, caffeine,chloroprocaine, choline, N,N′-dibenzyl-ethylenediamine (benzathine),dicyclohexylamine, diethanol-amine, diethyl-amine,2-diethyl-amino-ethanol, 2-dimethyl-amino-ethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethyl-piperidine, glucamine,glucosamine, histidine, hydrabamine, isopropyl-amine, lidocaine, lysine,meglumine (N-methyl-D-glucamine), morpholine, piperazine, piperidine,polyamine resins, procaine, purines, theobromine, triethanol-amine,triethylamine, trimethylamine, tripropyl-amine andtris(hydroxy-methyl)-methylamine (tromethamine), but this is notintended to represent a restriction.

Compounds of the formula (I) of the present invention which containbasic nitrogen-containing groups can be quaternised using agents such as(C1-C4)-alkyl halides, for example methyl, ethyl, isopropyl andtert-butyl chloride, bromide and iodide; di(C1-C4)alkyl sulfates, forexample dimethyl, diethyl and diamyl sulfate; (C10-C18)alkyl halides,for example decyl, do-decyl, lauryl, myristyl and stearyl chloride,bromide and iodide; and aryl-(C1-C4)alkyl halides, for example benzylchloride and phenethyl bromide. Both water- and oil-soluble compounds ofthe formula (I) can be prepared using such salts.

The above-mentioned pharmaceutical salts which are preferred includeacetate, trifluoroacetate, besylate, citrate, fumarate, gluconate,hemisuccinate, hippurate, hydrochloride, hydrobromide, isethionate,mandelate, meglumine, nitrate, oleate, phosphonate, pivalate, sodiumphosphate, stearate, sulfate, sulfosalicylate, tartrate, thiomalate,tosylate and tromethamine, but this is not intended to represent arestriction.

The acid-addition salts of basic compounds of the formula (I) areprepared by bringing the free base form into contact with a sufficientamount of the desired acid, causing the formation of the salt in aconventional manner. The free base can be regenerated by bringing thesalt form into contact with a base and isolating the free base in aconventional manner. The free base forms differ in a certain respectfrom the corresponding salt forms thereof with respect to certainphysical properties, such as solubility in polar solvents; for thepurposes of the invention, however, the salts other-wise correspond tothe respective free base forms thereof.

As mentioned, the pharmaceutically acceptable base-addition salts of thecompounds of the formula (I) are formed with metals or amines, such asalkali metals and alkaline earth metals or organic amines. Preferredmetals are sodium, potassium, magnesium and calcium. Preferred organicamines are N,N′-dibenzylethylenediamine, chloroprocaine, choline,diethanol-amine, ethylenediamine, N-methyl-D-glucamine and procaine.

The base-addition salts of acidic compounds of the formula (I) areprepared by bringing the free acid form into contact with a sufficientamount of the desired base, causing the formation of the salt in aconventional manner. The free acid can be regenerated by bringing thesalt form into contact with an acid and isolating the free acid in aconventional manner. The free acid forms differ in a certain respectfrom the corresponding salt forms thereof with respect to certainphysical properties, such as solubility in polar solvents; for thepurposes of the invention, however, the salts other-wise correspond tothe respective free acid forms thereof.

If a compound of the formula (I) contains more than one group which iscapable of forming pharmaceutically acceptable salts of this type, theformula (I) also encompasses multiple salts. Typical multiple salt formsinclude, for example, bitartrate, diacetate, difumarate, dimeglumine,di-phosphate, disodium and trihydrochloride, but this is not intended torepresent a restriction.

With regard to that stated above, it can be seen that the term“pharmaceutically acceptable salt” in the present connection is taken tomean an active ingredient which comprises a compound of the formula (I)in the form of one of its salts, in particular if this salt form impartsimproved pharmacokinetic properties on the active ingredient comparedwith the free form of the active ingredient or any other salt form ofthe active ingredient used earlier. The pharmaceutically acceptable saltform of the active ingredient can also provide this active ingredientfor the first time with a desired pharmacokinetic property which it didnot have earlier and can even have a positive influence on thepharmacodynamics of this active ingredient with respect to itstherapeutic efficacy in the body.

Owing to their molecular structure, the compounds of the formula (I) canbe chiral and can accordingly occur in various enantiomeric forms. Theycan therefore exist in racemic or in optically active form.

Since the pharmaceutical activity of the racemates or stereoisomers ofthe compounds according to the invention may differ, it may be desirableto use the enantiomers. In these cases, the end product or even theintermediates can be separated into enantiomeric compounds by chemicalor physical measures known to the person skilled in the art or evenemployed as such in the synthesis.

In the case of racemic amines, diastereomers are formed from the mixtureby reaction with an optically active resolving agent. Examples ofsuitable resolving agents are optically active acids, such as the R andS forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid,mandelic acid, malic acid, lactic acid, suitable N-protected amino acids(for example N-benzoylproline or N-benzenesulfonylproline), or thevarious optically active camphorsulfonic acids. Also advantageous ischromatographic enantiomer resolution with the aid of an opticallyactive resolving agent (for example dinitrobenzoylphenylglycine,cellulose triacetate or other derivatives of carbohydrates or chirallyderivatised methacrylate polymers immobilised on silica gel). Suitableeluents for this purpose are aqueous or alcoholic solvent mixtures, suchas, for example, hexane/isopropanol/acetonitrile, for example in theratio 82:15:3.

The invention furthermore relates to the use of compounds of formula(I), in combination with at least one further medicament activeingredient, preferably medicaments used in the treatment of multiplesclerosis such as cladribine or another co-agent, such as interferon,e.g. pegylated or non-pegylated interferons, preferably interferon betaand/or with compounds improving vascular function. These furthermedicaments, such as interferon beta, may be administered concomitantlyor sequentially, e.g. by subcutaneous, intramuscular or oral routes.These compositions can be used as medicaments in human and veterinarymedicine.

Pharmaceutical formulations can be administered in the form of dosageunits, which comprise a predetermined amount of active ingredient perdosage unit. Such a unit can comprise, for example, 0.5 mg to 1 g,preferably 1 mg to 700 mg, particularly preferably 5 mg to 100 mg, of acompound according to the invention, depending on the disease conditiontreated, the method of administration and the age, weight and conditionof the patient, or pharmaceutical formulations can be administered inthe form of dosage units which comprise a predetermined amount of activeingredient per dosage unit. Preferred dosage unit formulations are thosewhich comprise a daily dose or part-dose, as indicated above, or acorresponding fraction thereof of an active ingredient. Furthermore,pharmaceutical formulations of this type can be prepared using aprocess, which is generally known in the pharmaceutical art.

Pharmaceutical formulations can be adapted for administration via anydesired suitable method, for example by oral (including buccal orsublingual), rectal, nasal, topical (including buccal, sublingual ortransdermal), vaginal or parenteral (including subcutaneous,intramuscular, intravenous or intradermal) methods. Such formulationscan be prepared using all processes known in the pharmaceutical art by,for example, combining the active ingredient with the excipient(s) oradjuvant(s).

Pharmaceutical formulations adapted for oral administration can beadministered as separate units, such as, for example, capsules ortablets; powders or granules; solutions or suspensions in aqueous ornon-aqueous liquids; edible foams or foam foods; or oil-in-water liquidemulsions or water-in-oil liquid emulsions.

Thus, for example, in the case of oral administration in the form of atablet or capsule, the active-ingredient component can be combined withan oral, non-toxic and pharmaceutically acceptable inert excipient, suchas, for example, ethanol, glycerol, water and the like. Powders areprepared by comminuting the compound to a suitable fine size and mixingit with a pharmaceutical excipient comminuted in a similar manner, suchas, for example, an edible carbohydrate, such as, for example, starch ormannitol. A flavour, preservative, dispersant and dye may likewise bepresent.

Capsules are produced by preparing a powder mixture as described aboveand filling shaped gelatine shells therewith. Glidants and lubricants,such as, for example, highly disperse silicic acid, talc, magnesiumstearate, calcium stearate or polyethylene glycol in solid form, can beadded to the powder mixture before the filling operation. A disintegrantor solubiliser, such as, for example, agar-agar, calcium carbonate orsodium carbonate, may likewise be added in order to improve theavailability of the medicament after the capsule has been taken.

In addition, if desired or necessary, suitable binders, lubricants anddisintegrants as well as dyes can likewise be incorporated into themixture. Suitable binders include starch, gelatine, natural sugars, suchas, for example, glucose or beta-lactose, sweeteners made from maize,natural and synthetic rubber, such as, for example, acacia, tragacanthor sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes,and the like. The lubricants used in these dosage forms include sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, sodium chloride and the like. The disintegrants include,without being restricted thereto, starch, methylcellulose, agar,bentonite, xanthan gum and the like. The tablets are formulated by, forexample, preparing a powder mixture, granulating or dry-pressing themixture, adding a lubricant and a disintegrant and pressing the entiremixture to give tablets. A powder mixture is prepared by mixing thecompound comminuted in a suitable manner with a diluent or a base, asdescribed above, and optionally with a binder, such as, for example,carboxymethylcellulose, an alginate, gelatine or polyvinyl-pyrrolidone,a dissolution retardant, such as, for example, paraffin, an absorptionaccelerator, such as, for example, a quaternary salt, and/or anabsorbant, such as, for example, bentonite, kaolin or dicalciumphosphate. The powder mixture can be granulated by wetting it with abinder, such as, for example, syrup, starch paste, acadia mucilage orsolutions of cellulose or polymer materials and pressing it through asieve. As an alternative to granulation, the powder mixture can be runthrough a tableting machine, giving lumps of non-uniform shape which arebroken up to form granules. The granules can be lubricated by additionof stearic acid, a stearate salt, talc or mineral oil in order toprevent sticking to the tablet casting moulds. The lubricated mixture isthen pressed to give tablets. The active ingredients can also becombined with a free-flowing inert excipient and then pressed directlyto give tablets without carrying out the granulation or dry-pressingsteps. A transparent or opaque protective layer consisting of a shellacsealing layer, a layer of sugar or polymer material and a gloss layer ofwax may be present. Dyes can be added to these coatings in order to beable to differentiate between different dosage units.

Oral liquids, such as, for example, solution, syrups and elixirs, can beprepared in the form of dosage units so that a given quantity comprisesa pre-specified amount of the compounds. Syrups can be prepared bydissolving the compounds in an aqueous solution with a suitable flavour,while elixirs are prepared using a non-toxic alcoholic vehicle.Suspensions can be formulated by dispersion of the compounds in anon-toxic vehicle. Solubilisers and emulsifiers, such as, for example,ethoxylated isostearyl alcohols and polyoxyethylene sorbitol ethers,preservatives, flavour additives, such as, for example, peppermint oilor natural sweeteners or saccharin, or other artificial sweeteners andthe like, can likewise be added.

The dosage unit formulations for oral administration can, if desired, beencapsulated in microcapsules. The formulation can also be prepared insuch a way that the release is extended or retarded, such as, forexample, by coating or embedding of particulate material in polymers,wax and the like.

The compounds of the formula (I) and salts, solvates and physiologicallyfunctional derivatives thereof and the other active ingredients can alsobe administered in the form of liposome delivery systems, such as, forexample, small unilamellar vesicles, large unilamellar vesicles andmultilamellar vesicles. Liposomes can be formed from variousphospholipids, such as, for example, cholesterol, stearylamine orphosphatidylcholines.

The compounds of the formula (I) and the salts, solvates andphysiologically functional derivatives thereof and the other activeingredients can also be delivered using monoclonal antibodies asindividual carriers to which the compound molecules are coupled. Thecompounds can also be coupled to soluble polymers as targeted medicamentcarriers. Such polymers may encompass polyvinylpyrrolidone, pyrancopolymer, polyhydroxypropyl-methacrylamidophenol,polyhydroxyethylaspartamidophenol or polyethylene oxide polylysine,substituted by palmitoyl radicals. The compounds may furthermore becoupled to a class of biodegradable polymers which are suitable forachieving controlled release of a medicament, for example polylacticacid, poly-epsilon-caprolactone, polyhydroxybutyric acid,poly-orthoesters, polyacetals, polydihydroxypyrans, polycyanoacrylatesand crosslinked or amphipathic block copolymers of hydrogels.

Pharmaceutical formulations adapted for transdermal administration canbe administered as independent plasters for extended, close contact withthe epidermis of the recipient. Thus, for example, the active ingredientcan be delivered from the plaster by iontophoresis, as described ingeneral terms in Pharmaceutical Research, 3(6), 318 (1986).

Pharmaceutical compounds adapted for topical administration can beformulated as ointments, creams, suspensions, lotions, powders,solutions, pastes, gels, sprays, aerosols or oils.

For the treatment of the eye or other external tissue, for example mouthand skin, the formulations are preferably applied as topical ointment orcream. In the case of formulation to give an ointment, the activeingredient can be employed either with a paraffinic or a water-misciblecream base. Alternatively, the active ingredient can be formulated togive a cream with an oil-in-water cream base or a water-in-oil base.

Pharmaceutical formulations adapted for topical application to the eyeinclude eye drops, in which the active ingredient is dissolved orsuspended in a suitable carrier, in particular an aqueous solvent.

Pharmaceutical formulations adapted for topical application in the mouthencompass lozenges, pastilles and mouthwashes.

Pharmaceutical formulations adapted for rectal administration can beadministered in the form of suppositories or enemas.

Pharmaceutical formulations adapted for nasal administration in whichthe carrier substance is a solid comprise a coarse powder having aparticle size, for example, in the range 20-500 microns, which isadministered in the manner in which snuff is taken, i.e. by rapidinhalation via the nasal passages from a container containing the powderheld close to the nose. Suitable formulations for administration asnasal spray or nose drops with a liquid as carrier substance encompassactive-ingredient solutions in water or oil.

Pharmaceutical formulations adapted for administration by inhalationencompass finely particulate dusts or mists, which can be generated byvarious types of pressurised dispensers with aerosols, nebulisers orinsufflators.

Pharmaceutical formulations adapted for vaginal administration can beadministered as pessaries, tampons, creams, gels, pastes, foams or sprayformulations.

Pharmaceutical formulations adapted for parenteral administrationinclude aqueous and non-aqueous sterile injection solutions comprisingantioxidants, buffers, bacteriostatics and solutes, by means of whichthe formulation is rendered isotonic with the blood of the recipient tobe treated; and aqueous and non-aqueous sterile suspensions, which maycomprise suspension media and thickeners. The formulations can beadministered in single-dose or multidose containers, for example sealedampoules and vials, and stored in freeze-dried (lyophilised) state, sothat only the addition of the sterile carrier liquid, for example waterfor injection purposes, immediately before use is necessary.

Injection solutions and suspensions prepared in accordance with therecipe can be prepared from sterile powders, granules and tablets.

It goes without saying that, in addition to the above particularlymentioned constituents, the formulations may also comprise other agentsusual in the art with respect to the particular type of formulation;thus, for example, formulations which are suitable for oraladministration may comprise flavours.

A therapeutically effective amount of a compound of the formula (I) andof the other active ingredient depends on a number of factors,including, for example, the age and weight of the animal, the precisedisease condition which requires treatment, and its severity, the natureof the formulation and the method of administration, and is ultimatelydetermined by the treating doctor or vet. However, an effective amountof a compound is generally in the range from 0.1 to 100 mg/kg of bodyweight of the recipient (mammal) per day and particularly typically inthe range from 1 to 10 mg/kg of body weight per day. Thus, the actualamount per day for an adult mammal weighing 70 kg is usually between 70and 700 mg, where this amount can be administered as an individual doseper day or usually in a series of part-doses (such as, for example, two,three, four, five or six) per day, so that the total daily dose is thesame. An effective amount of a salt or solvate or of a physiologicallyfunctional derivative thereof can be determined as the fraction of theeffective amount of the compound per se.

The present invention furthermore relates to a method for treating asubject suffering from a sphingosine 1-phosphate associated disorder,comprising administering to said subject an effective amount of acompound of formula (I). The present invention preferably relates to amethod, wherein the sphingosine 1-phosphate-1 associated disorder is anautoimmune disorder or condition associated with an overactive immuneresponse.

The present invention furthermore relates to a method of treating asubject suffering from an immunerogulatory abnormality, comprisingadministering to said subject a compound of formula (I) in an amountthat is effective for treating said immunoregulatory abnormality. Thepresent invention preferably relates to a method wherein theimmunoregulatory abnormality is an autoimmune or chronic inflammatorydisease selected from the group consisting of: amyotrophic lateralsclerosis (ALS), systemic lupus erythematosus, chronic rheumatoidarthritis, type I diabetes mellitus, inflammatory bowel disease, biliarycirrhosis, uveitis, multiple sclerosis, Crohn's disease, ulcerativecolitis, bullous pemphigoid, sarcoidosis, psoriasis, autoimmunemyositis, Wegener's granulomatosis, ichthyosis, Graves opthalmopathy andasthma. The present invention furthermore relates to a method whereinthe immunoregulatory abnormality is bone marrow or organ transplantrejection or graft-versus-host disease. The present inventionfurthermore relates to a method wherein the immunoregulatory abnormalityis selected from the group consisting of: transplantation of organs ortissue, graft-versus-host diseases brought about by transplantation,autoimmune syndromes including rheumatoid arthritis, systemic lupuserythematosus, Hashimoto's thyroiditis, multiple sclerosis, myastheniagravis, type I diabetes, uveitis, posterior uveitis, allergicencephalomyelitis, glomerulonephritis, post-infectious autoimmunediseases including rheumatic fever and post-infectiousglomerulonephritis, inflammatory and hyperproliferative skin diseases,psoriasis, atopic dermatitis, contact dermatitis, eczematous dermatitis,seborrhoeic dermatitis, lichen planus, pemphigus, bullous pemphigoid,epidermolysis bullosa, urticaria, angioedemas, vasculitis, erythema,cutaneous eosinophilia, lupus erythematosus, acne, alopecia greata,keratoconjunctivitis, vernal conjunctivitis, uveitis associated withBehcet's disease, keratitis, herpetic keratitis, conical cornea,dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus,Mooren's ulcer, scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Haradasyndrome, sarcoidosis, pollen allergies, reversible obstructive airwaydisease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsicasthma, dust asthma, chronic or inveterate asthma, late asthma andairway hyper-responsiveness, bronchitis, gastric ulcers, vascular damagecaused by ischemic diseases and thrombosis, ischemic bowel diseases,inflammatory bowel diseases, necrotizing enterocolitis, intestinallesions associated with thermal burns, coeliac diseases, proctitis,eosinophilic gastroenteritis, mastocytosis, Crohn's disease, ulcerativecolitis, migraine, rhinitis, eczema, interstitial nephritis,Goodpasture's syndrome, hemolytic-uremic syndrome, diabetic nephropathy,multiple myositis, Guillain-Barre syndrome, Meniere's disease,polyneuritis, multiple neuritis, mononeuritis, radiculopathy,hyperthyroidism, Basedow's disease, pure red cell aplasia, aplasticanemia, hypoplastic anemia, idiopathic thrombocytopenic purpura,autoimmune hemolytic anemia, agranulocytosis, pernicious anemia,megaloblastic anemia, anerythroplasia, osteoporosis, sarcoidosis,fibroid lung, idiopathic interstitial pneumonia, dermatomyositis,leukoderma vulgaris, ichthyosis vulgaris, photoallergic sensitivity,cutaneous T cell lymphoma, chronic lymphocytic leukemia,arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritisnodosa, myocardosis, scleroderma, Wegener's granuloma, Sjogren'ssyndrome, adiposis, eosinophilic fascitis, lesions of gingiva,periodontium, alveolar bone, substantia ossea dentis,glomerulonephritis, male pattern alopecia or alopecia senilis bypreventing epilation or providing hair germination and/or promoting hairgeneration and hair growth, muscular dystrophy, pyoderma and Sezary'ssyndrome, Addison's disease, ischemia-reperfusion injury of organs whichoccurs upon preservation, transplantation or ischemic disease,endotoxin-shock, pseudomembranous colitis, colitis caused by drug orradiation, ischemic acute renal insufficiency, chronic renalinsufficiency, toxinosis caused by lung-oxygen or drugs, lung cancer,pulmonary emphysema, cataracta, siderosis, retinitis pigmentosa, senilemacular degeneration, vitreal scarring, corneal alkali burn, dermatitiserythema multiforme, linear IgA ballous dermatitis and cementdermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseasescaused by environmental pollution, aging, carcinogenesis, metastasis ofcarcinoma and hypobaropathy, disease caused by histamine orleukotriene-C₄ release, Behcet's disease, autoimmune hepatitis, primarybiliary cirrhosis, sclerosing cholangitis, partial liver resection,acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock,or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis,alcoholic cirrhosis, hepatic failure, fulminant hepatic failure,late-onset hepatic failure, “acute-on-chronic” liver failure,augmentation of chemotherapeutic effect, cytomegalovirus infection, HCMVinfection, AIDS, cancer, senile dementia, trauma, and chronic bacterialinfection.

Preferred compounds of formula (I) exhibit a binding constant Ki for thebinding to the S1P₁ receptor of less than about 5 μM, preferably lessthan about 1 μM and even more preferred less than about 0.1 μM.

Nomenclature of the compounds of this invention has been determinedusing ACD/Name Version 7.10 software.

In the following the present invention shall be illustrated by means ofsome examples, which are not construed to be viewed as limiting thescope of the invention.

EXAMPLES

The HPLC data provided in the examples described below were obtained asfollowed: Method A: HPLC columns: Xbridge™ C₈ column 50 mm×4.6 mm at aflow of 2 mL/min with 8 min gradient from 0.1% TFA in H₂O to 0.07% TFAin CH₃CN. UV detection (maxplot).

Method B: HPLC columns: BDS C₁₈ column 50 mm×4.6 mm at a flow of 0.8mL/min with 8 minutes gradient from 0.1% TFA in H₂O to CH₃CN. UVdetection (maxplot)

The MS data provided in the examples described below were obtained asfollowed: LC/MS Waters ZMD (ESI).

The NMR data provided in the examples described below were obtained asfollowed: ¹H-NMR: Bruker DPX-300 MHz or 400 MHz

The microwave chemistry is performed on a single mode microwave reactorEmrys™ Optimiser from Personal Chemistry.

General Procedure 1 for the Formation of Oxadiazole Derivatives

Trichloroacetonitrile was added to a suspension of the benzoic acid ofFormula (IV) and polymer bound triphenylphosphine in THF (2 mL) and thereaction mixture was stirred at 100° C. for 5 minutes in the microwave.A solution of the amidoxime of Formula (III) and DIEA in THF (2 mL) wasthen added and the reaction mixture was stirred at 150° C. for 15minutes in the microwave. The reaction mixture was then filtered througha SPE-NH₂ column, which was further washed with THF. After concentrationin vacuo, the residue was purified by column chromatography and/orcrystallization.

General Procedure 2 for the Formation of Oxadiazole Derivatives

Oxalyl chloride was added to a suspension of the benzoic acid of Formula(IV) and DMF (catalytic amount) in DCM (2 mL) and the reaction mixturewas stirred at room temperature for 30 minutes to 1 hour. Afterconcentration to dryness, the residue was taken up in THF (2 mL) andadded to a solution of the amidoxime of Formula (III) and DIEA in THF (1mL). The reaction mixture was then stirred at 150° C. for 30 minutes inthe microwave. After cooling, the mixture was filtered through a SPE-NH₂column, which was further washed with THF. After concentration in vacuo,the residue was purified by column chromatography and/orcrystallization.

Intermediate 1 N′-Hydroxy-2-methoxybenzenecarboximidamide

Hydroxylamine (Fluka 55458; 50% in water; 11.28 mL; 187.76 mmol; 5 eq.)was added to a solution of 2-methoxybenzonitrile (Alrich 231231; 4.59mL; 37.55 mmol; 1 eq.) in EtOH (50 mL) and the resulting mixture wasstirred at room temperature for 16 hours then at 55° C. for 24 hours.The solvent was then evaporated and the resulting colourless oil wasfurther dried under high vacuum to give a white solid. The latter wastriturated in n-hexane, filtered and dried to afford the title compound(6.29 g, quantitative) as a white solid.

HPLC (Method A): Rt 0.96 min (purity 99.2%).

Intermediate 2 N′-Hydroxy-2-(trifluoromethoxy)benzenecarboximidamide

Hydroxylamine (Fluka 55458; 50% in water; 4.81 mL; 80.16 mmol; 5 eq.)was added to a solution of 2-(trifluoromethoxy)benzonitrile (ApolloPC7438E; 3 g; 16.03 mmol; 1 eq.) in EtOH (20 mL) and the reactionmixture was stirred at 60° C. for 10 hours. Evaporation in vacuo gave awhite solid, which was further dried under high vacuum to afford thetitle compound (3.30 g, 94%) as a white solid.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.61 (1H, s), 760-7.47 (2H, m), 7.45-7.35(2H, m), 5.80 (2H, s).

Intermediate 3 2-Ethoxy-N′-hydroxybenzenecarboximidamide

Hydroxylamine (Fluka 55458; 50% in water; 4.08 mL; 67.95 mmol; 5 eq.)was added to a solution of 2-ethoxybenzonitrile (Fluorochem 18661; 2 g;13.59 mmol; 1 eq.) in EtOH (30 mL) and the reaction mixture was stirredat 50° C. for 12 hours. Evaporation in vacuo gave a white solid, whichwas further dried under high vacuum to afford the title compound (2.38g, 97%) as a white solid.

HPLC (Method A): Rt 1.22 min (purity 98.2%). LC/MS: 181.0 (M+H)⁺.

Intermediate 4 3-Nitro-4-piperidin-1-ylbenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.) and piperidine (Fluka 80640; 599.18 mg; 7.04 mmol; 3eq.) in DMF (2 mL) was stirred at 50° C. for 3 hours. The reaction wasthen allowed to cool to room temperature and diluted with water.Extraction with ethyl acetate, drying over sodium sulfate andconcentration in vacuo gave a yellow oil. The oil was taken up in THF(15 mL) and lithium hydroxide (280.86 mg; 11.73 mmol; 5 eq.) was added,followed by water (15 mL). The reaction mixture was then stirred at roomtemperature for 5 hours. THF was evaporated and the residue diluted withwater. The aqueous phase was washed with Et₂O and acidified to pH 5 withacetic acid. Extraction with Et₂O followed by drying over magnesiumsulfate and concentration in vacuo afforded the title compound (562 mg,96%) as a yellow solid.

HPLC (Method A): Rt 3.69 min (purity 99.7%). LC/MS: 250.9 (M−H)⁺, 252.9(M+H)⁺. ¹H NMR (DMSO-d₆, 300 MHz) δ 13.01 (1H, s), 8.23 (1H, s), 7.96(1H, d, J=8.3 Hz), 7.29 (1H, d, J=8.3 Hz), 3.09 (4H, s), 1.58 (6H, s).

Intermediate 5 4-Morpholin-4-yl-3-nitrobenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.) and morpholine (Fluka 69880; 613.06 mg; 7.04 mmol; 3eq.) in DMF (2 mL) was stirred at 50° C. for 3 hours. The reaction wasthen allowed to return to room temperature and diluted with water.Extraction with ethyl acetate, drying over sodium sulfate andconcentration in vacuo gave a yellow oil. The latter was taken up in THF(15 mL) and lithium hydroxide (280.86 mg; 11.73 mmol; 5 eq.) was addedfollowed by water (15 mL). The reaction mixture was stirred at roomtemperature for 5 hours. The THF was evaporated and the residue dilutedwith water. The aqueous phase was washed with Et₂O and acidified to pH 5with acetic acid. Extraction with Et₂O followed by drying over magnesiumsulfate and concentration in vacuo afforded the title compound (548 mg,93%) as a yellow solid.

¹H NMR (DMSO-d₆, 300 MHz) δ 13.12 (1H, s), 8.27 (1H, s), 8.02 (1H, d,J=8.3 Hz), 7.32 (1H, d, J=8.3 Hz), 3.72-3.65 (4H, m), 3.16-3.10 (4H, m).

Intermediate 6 4-Piperidin-1-ylbenzoic acid

A mixture of methyl 4-fluorobenzoate (Lancaster 14154; 500 mg; 3.24mmol; 1 eq.) and piperidine (Fluka 80640; 828.62 mg; 9.73 mmol; 3 eq.)in DMF (2 mL) was stirred at 50° C. for 16 hours. The reaction mixturewas then passed through a short pad of silica and the obtained solutionevaporated in vacuo to give a colourless oil. The oil was taken up inTHF (15 mL) and lithium hydroxide (388.41 mg; 16.22 mmol; 5 eq.) wasadded followed by water (15 mL). The reaction mixture was stirred atroom temperature for 6 hours then at 60° C. for 16 hours. NaOH (155.5mg, 6.48 mmol, 2 eq) was then added and the reaction mixture wasrefluxed for 24 hours. After cooling, the solution was diluted withwater and washed with Et₂O. The aqueous layer was then acidified to pH5-6 with acetic acid and the formed precipitate was collected byfiltration, washed with water and dried under high vacuum to afford thetitle compound as a white solid.

HPLC (Method A): Rt 1.43 min (purity 93.9%). LC/MS: 205.9 (M+H)⁺, 203.9(M−H)⁻.

Intermediate 7 4-[Cyclohexyl(methyl)amino]-3-nitrobenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.) and N-methylcyclohexylamine (Aldrich 10,332-2; 796.59mg; 7.04 mmol; 3 eq.) in DMF (2 mL) was stirred at 50° C. for 3 hours.The reaction was then allowed to return to room temperature and dilutedwith water. Extraction with ethyl acetate, drying over sodium sulfateand concentration in vacuo gave a yellow oil. The oil was taken up inTHF (15 mL) and lithium hydroxide (280.86 mg; 11.73 mmol; 5 eq.) wasadded followed by water (15 mL). The reaction mixture was stirred atroom temperature for 5 hours. After evaporation of the solvent, thesolution was diluted with water and washed with Et₂O. The aqueous layerwas acidified to pH 4 with AcOH, extracted with Et₂O, dried overmagnesium sulfate and concentrated in vacuo to afford the title compoundas a yellow solid.

HPLC (Method A): Rt 4.42 min (purity 98.3%). LC/MS: 277.0 (M−H)⁻.

Intermediate 8 4-(2-Methylpiperidin-1-yl)-3-nitrobenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 1.00 g;4.69 mmol; 1 eq.) and 2-methylpiperidine (Aldrich M7,280-3; 1.396 g;14.07 mmol; 3 eq.) in DMF (4 mL) was stirred at 50° C. for 3 hours. Thereaction was then allowed to return to room temperature and diluted withwater. Extraction with ethyl acetate, drying over sodium sulfate andconcentration in vacuo gave a yellow oil. The oil was taken up in THF(10 mL) and lithium hydroxide (561.73 mg; 23.46 mmol; 5 eq.) was addedfollowed by water (10 mL). The reaction mixture was stirred at roomtemperature for 16 hours. After evaporation of the THF, the solution wasdiluted with water and washed with Et₂O. The aqueous layer was acidifiedto pH 5 with AcOH, extracted with Et₂O, dried over magnesium sulfate andconcentrated in vacuo to afford the title compound (1.17 g, 94%) as ayellow solid.

LC/MS: 265.0 (M+H)⁺, 263.0 (M−H)⁻. ¹H NMR: (DMSO-d₆, 300 MHz) δ 13.07(s, 1H), 8.23 (d, J=2.1 Hz, 1H), 8.02 (dd, J=9.0, 2.3 Hz, 1H), 7.44-7.41(d, J=8.9 Hz, 1H), 3.64-3.60 (m, 1H), 3.25-3.17 (m, 1H), 2.90-2.84 (m,1H), 1.82-1.43 (m, 6H), 1.05 (d, J=6.4 Hz, 3H).

Intermediate 9 4-(3,3-Difluoropiperidin-1-yl)-3-nitrobenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.), 3,3-difluoropiperidine hydrochloride (Aldrich 665517;554.47 mg; 3.52 mmol; 1.5 eq.) and triethylamine (712.07 mg; 7.04 mmol;3 eq.) in DMF (3 mL) was stirred at 60° C. for 4 hours. The reactionmixture was then partitioned between ethyl acetate and aq. NH₄Cl. Theorganic layer was washed three times with aq. NH₄Cl then brine, driedover magnesium sulfate and concentrated in vacuo to give a yellow oil.The oil was taken up in THF (15 mL) and lithium hydroxide (280.86 mg;11.73 mmol; 5 eq.) then water (15 mL) were added. The reaction mixturewas stirred at room temperature for 4 hours and the THF was evaporatedunder vacuum. The resulting solution was washed with Et₂O, acidifiedwith acetic acid and extracted twice with Et₂O. The combined organiclayer was dried over magnesium sulfate and concentrated in vacuo to givea yellow oil which was crystallized from water. Filtration and dryingunder high vacuum afforded the title compound (630 mg, 94%) as a yellowsolid.

HPLC (Method A): Rt 3.61 min (purity 99.4%). LC/MS: 286.9 (M+H)⁺, 284.9(M−H)⁻.

Intermediate 10 3-Nitro-4-pyrrolidin-1-ylbenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.) and pyrrolidine (Fluka 83240; 500.48 mg; 7.04 mmol; 3eq.) in DMF (2 mL) was stirred at 60° C. for 4 hours. The reactionmixture was then partitioned between ethyl acetate and aq. NH₄Cl. Theorganic layer was washed three times with aq. NH₄Cl then brine, driedover magnesium sulfate and concentrated in vacuo to give a yellow oil.The latter was taken up in THF (5 mL) and lithium hydroxide (280.86 mg;11.73 mmol; 5 eq.) then water (5 mL) were added. The reaction mixturewas stirred at room temperature for 16 hours then the THF was evaporatedand the residue was diluted with water. After acidification with AcOH,the formed precipitate was filtered and dried under high vacuum toafford the title compound (497 mg, 90%) as a yellow solid.

HPLC (Method A): Rt 3.75 min (purity 99.0%). LC/MS: 236.8 (M+H)⁺, 234.9(M−H)⁻.

Intermediate 11 4-Azepan-1-yl-3-nitrobenzoic acid

A mixture of ethyl 4-fluoro-3-nitrobenzoate (Chontech 01072; 500 mg;2.35 mmol; 1 eq.) and hexamethyleneimine (Fluka 52660; 697.89 mg; 7.04mmol; 3 eq.) in DMF (2 mL) was stirred at 60° C. for 4 hours. Thereaction mixture was then partitioned between ethyl acetate and aq.NH₄Cl. The organic layer was washed with brine, dried over magnesiumsulfate and concentrated in vacuo to give a yellow solid. The latter wastaken up in THF (5 mL) and lithium hydroxide (280.86 mg; 11.73 mmol; 5eq.) then water (5 mL) were added. The reaction mixture was stirred atroom temperature for 16 hours then the solvent was evaporated and theresidue was diluted with water. After acidification to pH 5 with AcOH,the resulting precipitate was filtered and dried under high vacuum toafford the title compound (534 mg, 86%) as a yellow solid.

HPLC (Method A): Rt 3.16 min (purity 99.4%). LC/MS: 264.9 (M+H)⁺, 262.9(M−H)⁻.

Intermediate 12 4-Morpholin-4-yl-3-(trifluoromethyl)benzonitrile

A mixture of 4-fluoro-3-trifluoro-methylbenzonitrile (Fluorochem 2223;10 g; 52.8 mmol; 1 eq.) and morpholine (Fluka 69880; 9.25 mL; 105.7mmol; 2 eq.) was stirred at 60° C. for 8 hours. The mixture was cooledand diluted with water. The precipitate was filtered and dried to affordthe title compound (12.9 g, 95%) as a white solid.

HPLC (Method B): Rt 3.61 min (purity 99.1%). LC/MS: 257.1 (M+H)⁺. ¹H NMR(CDCl₃, 400 MHz) δ 7.92 (1H, s), 7.78-7.81 (1H, d), 7.32-7.35 (1H, d),3.84-3.87 (4H, m), 3.04-3.06 (4H, m).

Intermediate 13 Methyl 4-morpholin-4-yl-3-(trifluoromethyl)benzoate

A mixture of intermediate 12 (5 g, 19.5 mmol; 1 eq.) and HCl in Methanol(250 mL) was stirred at 60° C. for 24 hours. The reaction mixture wasevaporated to dryness and the residue partitioned between ethyl acetateand 10% aq. NaHCO₃. The separated organic layer was washed successivelywith water and brine, dried over magnesium sulfate and concentrated invacuo to afford the title compound (4.45 g, 97%) as a yellow oil.

¹H NMR (CDCl₃, 400 MHz) δ 8.32 (1H, s), 8.17-8.19 (1H, d), 7.31-7.33(1H, d), 3.94 (1H, s), 3.85-3.89 (4H, m), 3.02-3.07 (4H, m).

Intermediate 14 4-Morpholin-4-yl-3-(trifluoromethyl)benzoic acid

To a solution of intermediate 13 (5 g; 17.2 mmol; 1 eq.) in THF (50 mL)and water (5 mL) was added lithium hydroxide (1.5 g; 34.4 mmol; 2 eq.)and the reaction mixture was stirred at room temperature for 12 hours.The solvent was evaporated in vacuo and the residue diluted with water.This solution was washed with dichloromethane, acidified to pH 4 withconc. HCl and extracted with ethyl acetate. The organic layer was washedwith brine, dried over sodium sulfate and evaporated in vacuo to affordthe title compound as a white solid.

HPLC (Method B): Rt 2.97 min (purity 99.7%). LC/MS: 275.9 (M+H)⁺. ¹H NMR(DMSO-d₆, 400 MHz) δ 13.5 (1H, bs), 8.13-8.16 (2H, m), 7.55-7.57 (1H,d), 3.69-3.71 (4H, m), 2.94-2.96 (4H, m).

Intermediate 15 Methyl 2,2′-dimethyl-1,1′-biphenyl-4-carboxylate

A suspension of methyl 4-bromo-3-methylbenzoate (ABCR AV19078; 15 g;65.48 mmol; 1 eq.), o-tolylboronic acid (Aldrich 393606; 10.68 g; 78.5mmol; 1.2 eq.), potassium carbonate (45.25 g, 327.4 mmol, 5 eq.) andtetrakis(triphenylphosphine)palladium(0) (3.78 g; 3.27 mmol; 0.05 eq.)in toluene (200 mL) and water (200 mL) was stirred at 120° C. for 6hours. The resulting mixture was allowed to return to room temperatureand the two phases were separated. The organic layer was concentrated invacuo and purified by column chromatography (c-hexane) to afford thetitle compound (15 g, 95%) as a white solid.

HPLC (Method B): Rt 3.01 min (purity 98.7%). ¹H NMR (DMSO-d₆, 400 MHz) δ7.91 (1H, s), 7.83-7.81 (1H, m), 7.33-7.30 (2H, m), 7.28-7.26 (1H, m),7.25-7.22 (1H, m), 7.07-7.05 (1H, d), 3.86-3.81 (3H, s), 2.09-2.00 (3H,s), 1.97-1.92 (3H, s).

Intermediate 16 2,2′-Dimethyl-1,1′-biphenyl-4-carboxylic acid

Sodium hydroxide (10% in water; 10 mL) was added to a solution ofintermediate 15 (15 g, 62.24 mmol; 1 eq.) in THF (100 mL) and thereaction mixture was stirred at 70° C. for 16 hours. The solvent wasevaporated in vacuo and the aqueous residue washed with ethyl acetate.The aqueous layer was then acidified pH 2-3 with 3M HCl and extractedwith dichloromethane. The organic layer was washed with water, driedover sodium sulfate and concentrated in vacuo to afford the titlecompound (13.5 g, 95%) as a white solid.

HPLC (Method B): Rt 4.10 min (purity 99.6%). LC/MS: 227.0 (M+H)⁺. ¹H NMR(DMSO, 400 MHz) δ 12.89 (1H, bs), 7.89 (1H, s), 7.82-7.80 (1H, d),7.32-7.23 (3H, m), 7.19-7.11 (1H, d), 7.07-7.05 (1H, d), 2.04 (3H, s),1.98 (3H, s).

Intermediate 17 Methyl2-methyl-2′-(trifluoromethyl)biphenyl-4-carboxylate

A suspension of methyl 4-bromo-3-methylbenzoate (ABCR AV19078; 3 g;13.10 mmol; 1 eq.), 2-(trifluoromethyl)phenylboronic acid (Aldrich393606; 2.74 g; 14.41 mmol; 1.10 eq.), potassium carbonate (9.05 g;65.48 mmol; 5 eq.) and tetrakis(triphenylphosphine)palladium(0) (1.51 g;1.31 mmol; 0.10 eq.) in toluene (15 mL) and water (15 mL) was refluxedfor 3 hours. The resulting mixture was filtered through a short pad ofCelite, which was further washed with toluene. After evaporation of thesolvent, the residue was taken up in ethyl acetate and washedsuccessively with sat. aq. NaHCO₃, water and brine, dried over magnesiumsulfate and concentrated in vacuo to afford the title compound (3.7 g,96%) as a brown oil.

HPLC (Method A): Rt 5.34 min (purity 70.9%). LC/MS: 294.9 (M+H)⁺.

Intermediate 18 2-Methyl-2′-(trifluoromethyl)biphenyl-4-carboxylic acid

Sodium hydroxide (6.12 mL; 5M; 30.58 mmol; 3 eq.) was added to asolution of intermediate 17 (3 g; 10.19 mmol; 1 eq.) in EtOH (90 mL) andthe resulting mixture was stirred at 60° C. for 1 hour. Afterevaporation of the solvent, the residue was taken up in water and washedwith ethyl acetate. The aqueous phase was acidified to pH 2 with conc.HCl and the obtained solution was concentrated in vacuo untilcrystallization. The solid was collected by filtration and dried underhigh vacuum to afford the title compound (2.41 g, 84%) as a beige solid.

HPLC (Method A): Rt 4.49 min (purity 95.7%). LC/MS: 279.0 (M−H)⁻. ¹H NMR(DMSO-d₆, 300 MHz) δ13.03 (s, 1H), 7.91-7.68 (m, 5H), 7.38-7.36 (d,J=8.3 Hz, 1H), 7.27-7.25 (d, J=8.2 Hz, 1H), 2.05 (s, 3H).

Intermediate 19 Methyl 3-methyl-4-(4-methyl-3-thienyl)benzoate

A suspension of methyl 4-bromo-3-methylbenzoate (ABCR AV19078; 3 g;13.10 mmol; 1 eq.), 4-methyl-3-thiopheneboronic acid (Aldrich 542393;2.05 g; 14.41 mmol; 1.10 eq.), potassium carbonate (9.05 g; 65.48 mmol;5 eq.) and tetrakis(triphenylphosphine)palladium(0) (1.51 g; 1.31 mmol;0.10 eq.) in toluene (15 mL) and water (15 mL) was refluxed for 3 hours.The resulting mixture was filtered through a short pad of Celite, whichwas further washed with toluene. After evaporation of the solvent, theresidue taken up in ethyl acetate and washed successively with sat. aq.NaHCO₃, water and brine, dried over magnesium sulfate and concentratedin vacuo to afford the title compound (2.96 g, 92%) as a brown oil.

HPLC (Method A): Rt 5.14 min (purity 58.3%). LC/MS: 246.8 (M+H)⁺.

Intermediate 20 3-Methyl-4-(4-methyl-3-thienyl)benzoic acid

Sodium hydroxide (4.87 mL; 5M; 24.36 mmol; 3 eq.) was added to asolution of intermediate 19 (2 g; 8.12 mmol; 1 eq.) in EtOH (60 mL) andthe resulting mixture was stirred at 60° C. for 2 hours. Afterevaporation of the solvent, the residue was taken up in water and washedwith ethyl acetate. The aqueous phase was acidified to pH 2 with conc.HCl and the obtained solution was concentrated in vacuo untilcrystallization. The solid was collected by filtration and dried underhigh vacuum to afford the title compound as a beige solid.

HPLC (Method A): Rt 4.26 min (purity 99.6%). LC/MS: 232.9 (M+H)⁺, 231.0(M−H)⁻. ¹H NMR: (DMSO-d₆, 300 MHz) δ 12.97 (s, 1H), 7.91 (s, 1H),7.84-7.81 (dd, J=8.1 Hz, J=1.9 Hz; 1H), 7.41 (d, J=3.2 Hz, 1H),7.35-7.34 (m, 1H), 7.28-7.26 (d, J=7.8 Hz, 1H), 2.18 (s, 3H), 2.00 (s,3H).

Intermediate 21 Methyl 2′-methyl-2-nitro-1,1′-biphenyl-4-carboxylate

A suspension of methyl 4-bromo-3-nitrobenzoate (Chess 1687; 3 g; 11.53mmol; 1 eq.), o-tolylboronic acid (Aldrich 393606; 1.88 g; 13.84 mmol;1.2 eq.), potassium carbonate (7.97 g, 57.68 mmol, 5 eq.) andtetrakis(triphenylphosphine)palladium(0) (668.2 mg, 0.577 mmol, 0.05eq.) in toluene (15 mL) and water (15 mL) was stirred at 120° C. for 14hours. The resulting mixture was allowed to return to room temperatureand the two phases were separated. The organic layer was concentrated invacuo and purified by column chromatography (c-hexane) to afford thetitle compound (2.5 g, 79%) as a white solid.

¹H NMR (DMSO-d₆, 400 MHz) δ 8.52 (1H, s), 8.29-8.26 (1H, m), 7.63-7.61(1H, m), 7.34-7.33 (2H, m), 7.23 (1H, m), 7.13 (1H, m), 3.94 (3H, s),2.04 (3H, s).

Intermediate 22 2′-Methyl-2-nitro-1,1′-biphenyl-4-carboxylic acid

Lithium hydroxide (1.15 g, 27.6 mmol, 3 eq.) was added to a solution ofintermediate 21 (2.5 g, 9.2 mmol, 1 eq.) in THF (20 mL) and water (5 mL)and the reaction mixture was stirred at room temperature for 4 hours.The solvent was evaporated in vacuo and the aqueous residue washed withethyl acetate. The aqueous layer was then acidified pH 2-3 with 1.5M HCland extracted with dichloromethane. The organic layer was washed withwater, dried over sodium sulfate and concentrated in vacuo to afford thetitle compound (1.6 g, 70%) as a yellow solid. HPLC (Method B): Rt 3.70min (purity 99.9%). LCMS: 255.9 (M−H)⁻. ¹H NMR (DMSO-d₆, 400 MHz) δ13.67 (1H, bs), 8.49 (1H, s), 8.26-8.24 (1H, d), 7.58-7.56 (1H, m),7.35-7.32 (2H, m), 7.27-7.12 (1H, d), 7.10 (1H, d), 2.09-1.99 (3H, s).

Intermediate 23 5-fluoro-N′-hydroxy-2-methoxybenzenecarboximidamide

Hydroxylamine (4.97 ml; 82.7 mmol; 5 eq.) was added to a suspension of5-fluoro-2-methoxybenzonitrile (Aldrich 527734; 2.5 g; 16.5 mmol; 1 eq.)in EtOH (30 mL) and the resulting mixture was stirred at roomtemperature for 3 days then to dryness to afford the title compound(3.04 g, 100%) as a white solid.

¹H NMR (DMSO-d₆, 300 MHz) δ 9.53 (s, 1H), 7.25-7.15 (m, 2H), 7.11-7.03(m, 1H), 5.69 (s, 2H), 3.78 (s, 3H).

Intermediate 24 4-(2-Methylpiperidin-1-yl)-3-(trifluoromethyl)benzoicacid Step 1: 4-(2-Methylpiperidin-1-yl)-3-(trifluoromethyl)benzonitrile

A mixture of 4-fluoro 3-trifluoro-methylbenzonitrile (ABCR F043738; 25g; 132 mmol; 1 eq.) and 2-methylpiperidine (30.3 mL; 264 mmol; 2 eq.)was stirred at 100° C. 12 hours. The reaction mixture was cooled,diluted with water (200 mL) and extracted with ethyl acetate (2×200 mL).The combined organic layer was washed with water (100 mL) and brine (100mL), dried over sodium sulphate and concentrated in vacuo. The residuewas purified by column chromatography (petroleum ether/ethyl acetate,80/20) to afford the title compound of the title compound as anoff-white solid.

LC/MS: 269 (M+H)⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 8.19 (s, 1H), 8.18-8.15(m, 1H), 7.90-7.80 (m, 1H), 3.19-3.15 (m, 1H), 2.90-2.85 (m, 1H),2.71-2.60 (m, 1H), 1.81-1.75 (m, 2H), 1.60-1.31 (m, 3H), 1.31-1.21 (m,1H), 0.70 (d, J=6.4 Hz, 3H).

Step 2: Methyl 4-(2-methylpiperidin-1-yl)-3-(trifluoromethyl)benzoate.Hydrochloride salt

A mixture of 4-(2-methylpiperidin-1-yl)-3-(trifluoromethyl)benzonitrile(21 g, 78.4 mmol) and HCl in methanol (4M, 500 mL) was stirred at 80° C.for 20 hours, then concentrated to dryness to afford the title compound(25 g, 96%) as a beige solid.

LC/MS: 301.9 (M+H)⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 8.20-8.18 (m, 2H),7.69-7.62 (m, 1H), 3.89 (s, 3H), 3.20-3.12 (m, 1H), 2.90-2.85 (m, 1H),2.60-2.50 (m, 1H), 1.85-1.79 (m, 2H), 1.69-1.65 (m, 2H), 1.51-1.45 (m,1H), 1.31-1.23 (m, 1H), 0.75 (d, J=6.4 Hz, 3H).

Step 3: 4-(2-methylpiperidin-1-yl)-3-(trifluoromethyl)benzoic acid

A solution of methyl4-(2-methylpiperidin-1-yl)-3-(trifluoromethyl)benzoate. Hydrochloridesalt (25 g, 74 mmol) in HCl 4M (200 mL) was stirred at 100° C. for 16hours. After cooling to room temperature, the precipitate was filteredoff and dried under vacuum to afford the title compound (18 g, 85%) ofas a white solid.

HPLC (Method B): Rt 5.53 min (purity 98.3%). LC/MS: 287.9 (M+H)⁺. ¹H NMR(DMSO-d₆, 400 MHz) δ 13.30 (s, 1H), 8.19-8.15 (m, 2H), 7.70-7.67 (m,1H), 3.08-3.06 (m, 1H), 2.89-2.86 (m, 1H), 2.60-2.49 (m, 1H), 1.90-1.89(m, 2H), 1.76-1.74 (m, 2H), 1.46-1.43 (m, 1H), 1.41-1.40 (m, 1H), 0.77(d, J=6.4 Hz, 3H).

Intermediate 25 4-(2,5-dimethylpyrrolidin-1-yl)-3-nitrobenzoic acid

A mixture of 4-fluoro-3-nitrobenzoic acid (300 mg; 1.62 mmol; 1 eq.) and2,5-dimethylpyrrolidine (Maybridge AC11676DA; 0.70 mL; 5.72 mmol; 3.53eq.) was stirred at 100° C. for 5 minutes in the MW. The solution wasconcentrated in vacuo and the residue taken up in water (100 mL). Theaqueous phase was washed with diethyl ether (2×20 mL) then acidified topH 5 with acetic acid. Extraction with diethyl ether (150 mL), dryingover magnesium sulfate and concentration in vacuo afforded the titlecompound (360 mg, 84%) as an orange solid.

HPLC (Method A): Rt 3.82 min (purity 100%). LC/MS: 264.9 (M+H)⁺.

Intermediate 26 2′-methoxy-2-methylbiphenyl-4-carboxylic acid

Step 1: methyl 2′-methoxy-2-methylbiphenyl-4-carboxylate

A suspension of methyl 4-bromo-3-methylbenzoate (4.9 g; 21.4 mmol; 1eq.), 2-methoxyphenylboronic acid (Aldrich 44,523-1; 3.6 g; 23.5 mmol;1.1 eq.), potassium carbonate (14.8 g; 107 mmol; 5 eq.) and Pd(PPh₃)₄(2.47 g; 2.14 mmol; 0.1 eq.) in toluene (24.5 mL) and water (24.5 mL)was refluxed for 6 hours. The reaction mixture was cooled down to roomtemperature and filtered through a pad of Celite® which was furtherwashed with toluene (500 mL). The filtrate was concentrated in vacuo andthe residue taken up in ethyl acetate (500 mL). The organic layer waswashed with sat. aq. NaHCO₃ (200 mL), water (200 mL) and brine (200 mL)then dried over magnesium sulfate and concentrated in vacuo. The crudewas purified by column chromatography (c-hexane/ethyl acetate, 90/10) toafford the title compound (4.38 g, 80%) as a colorless oil. HPLC (MethodA): Rt 4.85 min (purity 98.9%). LC/MS: 257.0 (M+H)⁺.

Step 2: 2′-methoxy-2-methylbiphenyl-4-carboxylic acid

Sodium hydroxide (5M; 4.7 mL; 23.4 mmol; 3 eq.) was added to a solutionof methyl 2′-methoxy-2-methylbiphenyl-4-carboxylate (2 g; 7.8 mmol; 1eq.) in EtOH (60 mL) and the reaction mixture was stirred at 60° C. forone hour then concentrated in vacuo. The residue was taken up in water(400 mL) and washed with ethyl acetate (2×200 mL) then acidified to pH 2with conc. HCl. The solution was concentrated to ca. 80 mL, theprecipitate filtered off and dried to afford the title compound as abrown solid.

HPLC (Method A): Rt 4.05 min (purity 98.5%). LC/MS: 240.9 (M−H)⁻.

Intermediate 27 2-methoxy-2′-methylbiphenyl-4-carboxylic acid

A suspension of methyl 4-bromo-3-methoxybenzoate (Combi-blocks CA-4192;2.5 g; 10.2 mmol; 1 eq.), o-tolylboronic acid (Aldrich 393606; 1.53 g;11.2 mmol; 1.1 eq.), potassium carbonate (7.05 g; 51 mmol; 5 eq.) andPd(PPh₃)₄ (1.18 g; 1.02 mmol; 0.1 eq.) in toluene (12.5 mL) and water(12.5 mL) was refluxed for 2 hours. The reaction mixture was cooled downto room temperature and filtered through a pad of Celite® which wasfurther washed with toluene (200 mL). The filtrate was concentrated invacuo and the residue taken up in Ethyl acetate (500 mL). The organiclayer was washed with sat. aq. NaHCO₃ (150 mL), water (150 mL) and brine(150 mL) then dried over magnesium sulfate and concentrated in vacuo.The residue (2.5 g; 9.75 mmol; 1 eq.) was taken up in EtOH (75 mL),sodium hydroxide (5M; 5.85 mL; 29.3 mmol; 3 eq.) was added and thereaction mixture was stirred at 60° C. for two hours. Afterconcentration in vacuo, the residue was taken up in water (400 mL) andthe aqueous phase was washed with Ethyl acetate then acidified to pH 2with conc. HCl. The solution was concentrated to ca. 80 mL and theprecipitate filtered off and dried to afford the title compound (1.95 g,79%) as a beige solid.

HPLC (Method A): Rt 4.05 min (purity 97.3%). LC/MS: 240.9 (M−H)⁻.

Intermediate 28 2′,4′-dimethoxy-2-methylbiphenyl-4-carboxylic acid

A suspension of methyl 4-bromo-3-methylbenzoate (3.0 g; 13.1 mmol; 1eq.), 2,4-dimethoxyphenylboronic acid (Aldrich 483-486; 2.62 g; 14.4mmol; 1.1 eq.), potassium carbonate (9.05 g; 65.5 mmol; 5 eq.) andPd(PPh₃)₄ (1.51 g; 1.31 mmol; 0.1 eq.) in toluene (15 mL) and water (15mL) was refluxed for 6 hours. The reaction mixture was cooled down toroom temperature and filtered through a pad of Celite® which was furtherwashed with toluene (200 mL). The filtrate was concentrated in vacuo andthe residue taken up in Ethyl acetate (300 mL). The organic layer waswashed with sat. aq. NaHCO₃ (100 mL), water (100 mL) and brine (100 mL)then dried over magnesium sulfate and concentrated in vacuo. The residue(2.0 g; 6.99 mmol; 1 eq.) was taken up in EtOH (60 mL), sodium hydroxide(5M; 4.19 mL; 21 mmol; 3 eq.) was added and the reaction mixture wasstirred at 60° C. for one hour. After concentration in vacuo, theresidue was taken up in water (400 mL) and the aqueous phase was washedwith Ethyl acetate then acidified to pH 2 with conc. HCl. The solutionwas concentrated to ca. 80 mL and the precipitate filtered off and driedto afford the title compound as an orange solid.

HPLC (Method A): Rt 3.87 min (purity 99.7%). LC/MS: 270.9 (M−H)⁻.

Intermediate 29 3-methoxy-4-(4-methyl-3-thienyl)benzoic acid

A suspension of methyl 4-bromo-3-methoxybenzoate (2.5 g; 10.2 mmol; 1.00eq.), 4-methyl-3-thiopheneboronic acid (Aldrich 542393; 1.59 g; 11.2mmol; 1.1 eq.), potassium carbonate (7.05 g; 51 mmol; 5 eq.) andPd(PPh₃)₄ (1.18 g; 1 mmol; 0.1 eq.) in toluene (12.5 mL) and water (12.5mL) was refluxed for 5 hours. The reaction mixture was cooled down toroom temperature and filtered through a pad of Celite® which was furtherwashed with toluene (200 mL). The filtrate was concentrated in vacuo andthe residue taken up in Ethyl acetate (400 mL). The organic layer waswashed with sat. aq. NaHCO₃ (100 mL), water (100 mL) and brine (100 mL)then dried over magnesium sulfate and concentrated in vacuo. The residue(2.3 g; 8.77 mmol; 1 eq.) was taken up in EtOH (69 mL), sodium hydroxide(5M; 5.26 mL; 26.3 mmol; 3 eq.) was added and the reaction mixture wasstirred at 60° C. for one hour. After concentration in vacuo, theresidue was taken up in water (200 mL) and the aqueous phase was washedwith Ethyl acetate then acidified to pH 2 with conc. HCl. The solutionwas concentrated to ca. 100 mL and the precipitate filtered off anddried to afford the title compound (1.81 g, 71%) as a brown solid.

HPLC (Method A): Rt 3.99 min (purity 97.4%). LC/MS: 246.9 (M−H)⁻.

Intermediate 30 4-(3,5-dimethylisoxazol-4-yl)-3-methylbenzoic acid

A suspension of methyl 4-bromo-3-methylbenzoate (4 g; 17.5 mmol; 1 eq.),3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)isoxazole(Fluorochem 11035; 4.28 g; 19.2 mmol; 1.1 eq.), potassium carbonate(12.1 g; 87.3 mmol; 5 eq.) and Pd(PPh₃)₄ (2.02 g; 1.75 mmol; 0.1 eq.) intoluene (20 mL) and water (20 mL) was refluxed for 4 hours. The reactionmixture was cooled down to room temperature and filtered through a padof Celite® which was further washed with toluene (200 mL). The filtratewas concentrated in vacuo and the residue taken up in Ethyl acetate (250mL). The organic layer was washed with sat. aq. NaHCO₃ (100 mL), water(100 mL) and brine (100 mL) then dried over magnesium sulfate andconcentrated in vacuo. The residue (3 g; 12.2 mmol; 1 eq.) was taken upin EtOH (90 mL), sodium hydroxide (5M; 7.34 mL; 36.7 mmol; 3 eq.) wasadded and the reaction mixture was stirred at 60° C. for one hour. Afterconcentration in vacuo, the residue was taken up in water (200 mL) andthe aqueous phase was washed with Ethyl acetate then acidified to pH 2with conc. HCl. The solution was concentrated to ca. 100 mL and theprecipitate filtered off and dried. Trituration in Et₂O and filtrationafforded the title compound as a beige solid.

HPLC (Method A): Rt 3.13 min (purity 97.1%).

Intermediate 31 4-[bis(2-methoxyethyl)amino]-3-nitrobenzoic acid

Bis(2-methoxyethyl)amine (Aldrich B4,820-7; 1.6 g; 12 mmol; 3 eq.) wasadded to a solution of 4-fluoro-3-nitrobenzoic acid (740 mg; 4 mmol; 1eq.) in EtOH (20 mL) and the resulting mixture was stirred at roomtemperature for 2 hours then at 70° C. for 20 hours. The solution wasconcentrated in vacuo and the residue was partitioned between NaOH 0.5Mand Et₂O. The aqueous layer was acidified to pH 2 with HCl 5M, extractedwith ethyl acetate (3×) and the combined organic phase was dried overmagnesium sulfate. After evaporation of the solvent, the residue wascrystallised from ethyl acetate/n-pentane to afford the title compound(1.12 g, 94%) as a yellow solid.

¹H NMR (DMSO-d₆, 300 MHz) δ 12.95 (s, 1H), 8.17 (d, J=2.2 Hz, 1H), 7.93(dd, J=8.9, 2.2 Hz, 1H), 7.39 (d, J=8.9 Hz, 1H) 3.50-3.36 (m, 8H), 3.18(s, 6H).

Intermediate 32 4-(4-methyl-3-thienyl)-3-(trifluoromethyl)benzoic acid

Step 1: methyl 4-bromo-3-(trifluoromethyl)benzoate

Thionyl chloride (16.2 mL; 223 mmol; 4 eq.) was added to a suspension of4-bromo-3-(trifluoromethyl)benzoic acid (15 g; 55.8 mmol; 1 eq.) in MeOH(300 mL) and the reaction mixture was stirred at room temperature for 16hours.

The solvent was concentrated in vacuo and the residue was diluted withethyl acetate (500 mL). The organic layer was washed with sat. aq.NaHCO₃ (200 mL), water (200 mL) and brine (200 mL) then dried overmagnesium sulphate and concentrated in vacuo to afford the titlecompound (14.8 g, 94%) as an orange solid.

HPLC (Method A): Rt 4.71 min (purity 99.0%).

Step 2: methyl 4-(4-methyl-3-thienyl)-3-(trifluoromethyl)benzoate

A mixture of methyl 4-bromo-3-(trifluoromethyl)benzoate (3.5 g; 12.4mmol; 1 eq.), 4-methyl-3-thiopheneboronic acid (1.93 g; 13.6 mmol; 1.1eq.), potassium carbonate (8.54 g; 61.8 mmol; 5 eq.) and Pd(PPh₃)₄ (1.43g; 1.24 mmol; 0.1 eq.) in toluene (17.5 mL) and water (17.5 mL) wasrefluxed for 24 h whereupon 4-methyl-3-thiopheneboronic acid (0.88 g;6.2 mmol; 0.5 eq.) was added. The reaction mixture was stirred for afurther 3 hours then cooled down to room temperature and filteredthrough a pad of Celite® which was washed with toluene (200 mL). Thefiltrate was concentrated in vacuo and the residue taken up in ethylacetate (300 mL). The organic layer was washed with sat. aq. NaHCO₃ (100mL), water (100 mL) and brine (100 mL), dried over magnesium sulphateand concentrated in vacuo to afford the title compound (3 g, 81%) as abrown oil.

HPLC (Method A): Rt 5.21 min (purity 68.1%).

Step 3: 4-(4-methyl-3-thienyl)-3-(trifluoromethyl)benzoic acid

Sodium hydroxide (5M: 6 mL; 30 mmol; 3 eq.) was added to a solution ofmethyl 4-(4-methyl-3-thienyl)-3-(trifluoromethyl)benzoate (3 g; 10 mmol;1 eq.) in EtOH (90 mL) and the reaction mixture was stirred at 60° C.for 2 hours. After evaporation of the solvent, the residue was taken upin water (300 mL) and the aqueous phase was washed with ethyl acetate(2×100 mL) then acidified to pH 2 with conc. HCl. The solution wasconcentrated to ca. 100 mL and extracted with ethyl acetate. The organiclayer was dried over magnesium sulphate and concentrated in vacuo toafford the title compound (2.6 g, 91%) as a brown oil.

HPLC (Method A): Rt 4.53 min (purity 95.5%). LC/MS: 284.9 (M−H)⁻.

Intermediate 33 3-Methyl-4-piperidin-1-ylbenzoic acid

Step 1: Methyl 3-methyl-4-piperidin-1-ylbenzoate

BINAP (0.67 g; 1.1 mmol; 0.05 eq.) and palladium acetate (0.24 g; 1.1mmol; 0.05 eq.) were added to a suspension methyl4-bromo-3-methylbenzoate (5 g; 21.8 mmol; 1 eq.), Cs₂CO₃ (10.65 g; 32.7mmol; 1.5 eq.) and piperidine (2.2 g; 26 mmol; 1.2 eq.) in dioxane (100mL) and the reaction mixture was refluxed for 15 hours. After filtrationthrough a pad of Celite®, the solution was concentrated in vacuo and theresidue purified by column chromatography (petroleum ether/ethylacetate, 80/20) to afford the title compound (4.9 g, 96%) as a brownsolid.

LC/MS: 233.9 (M−H)⁺. ¹H NMR (CDCl₃, 400 MHz) δ 7.84-7.82 (m, 2H),6.99-6.97 (m, 1H), 3.92 (s, 3H), 2.93-2.90 (m, 4H), 1.76-1.67 (m, 4H,m), 1.63-1.62 (m, 2H).

Step 2: 3-Methyl-4-piperidin-1-ylbenzoic acid

Lithium hydroxide (2.5 g; 103 mmol; 5 eq.) was added to a solution ofmethyl 3-methyl-4-piperidin-1-ylbenzoate (4.8 g; 20.6 mmol; 1 eq.) inTHF (100 mL) and water (5 mL) and the reaction mixture was stirred at50° C. for 12 hours. The solvent was removed in vacuo, the residuediluted with water and the aqueous layer washed with DCM (2×50 mL). Theaqueous layer was acidified to pH 4 with conc. HCl and extracted withethyl acetate (2×100 mL). The combined organic layer was washed withbrine, dried over sodium sulfate and concentrated in vacuo to afford thetitle compound (4.0 g, 89%) as a yellow solid.

HPLC (Method B): Rt 5.47 min (purity 99.0%). LC/MS: 219.9 (M+H)⁺.

¹H NMR (DMSO-d₆, 400 MHz) δ 7.71-7.70 (m, 2H), 7.01-6.99 (m, 1H),2.84-2.82 (m, 4H), 2.25 (s, 3H), 1.64-1.59 (m, 4H), 1.54-1.53 (m, 2H).

Intermediate 34 2′methyl-2-(trifluoromethyl)biphenyl-4-carboxylic acid

Step 1: methyl 2′-methyl-2-(trifluoromethyl)biphenyl-4-carboxylate

A mixture of methyl 4-bromo-3-(trifluoromethyl)benzoate (6 g; 21.2 mmol;1 eq.), o-tolylboronic acid (3.17 g; 23.3 mmol; 1.1 eq.), potassiumcarbonate (14.65 g; 106 mmol; 5 eq.) and Pd(PPh₃)₄ (2.45 g; 2.12 mmol;0.1 eq.) in toluene (30 mL) and water (30 mL) was refluxed for 3 hours.After cooling to room temperature, the reaction mixture was filteredthrough a pad of Celite® which was further washed with toluene (20 mL).The filtrate was concentrated in vacuo, the residue taken up in ethylacetate (200 mL) and washed with sat. aq. NaHCO₃ (100 mL), water (100mL) and brine (100 mL), dried over magnesium sulphate and concentratedin vacuo to afford the title compound (5 g, 80%) as a brown oil

HPLC (Method A): Rt 5.33 min (purity 60.0%).

Step 2: 2′-methyl-2-(trifluoromethyl)biphenyl-4-carboxylic acid

Sodium hydroxide (5M; 10.2 mL; 51 mmol; 3 eq.) was added to a solutionof methyl 2′-methyl-2-(trifluoromethyl)biphenyl-4-carboxylate (5 g; 17mmol; 1 eq.) in EtOH (150 mL) and the reaction mixture was stirred at60° C. for 2 hours. After concentration in vacuo, the residue was takenup in water (300 mL) and the aqueous phase was washed with ethyl acetate(2×100 mL). The pH was adjusted to 2 with conc. HCl and the solution wasconcentrated to ca. 150 mL. The precipitate was filtered off and driedto afford the title compound (3.33 g, 70%) as a beige solid. HPLC(Method A): Rt 4.57 min (purity 98.7%). LC/MS: 278.9 (M−H)⁻.

Intermediate 35 3-Methyl-4-morpholin-4-ylbenzoic acid

Step 1: Methyl 3-methyl-4-morpholin-4-ylbenzoate

BINAP (0.67 g; 1.1 mmol; 0.05 eq.) and palladium acetate (0.24 g; 1.1mmol; 0.05 eq.) were added to a suspension methyl4-bromo-3-methylbenzoate (5 g; 21.8 mmol; 1 eq.), Cs₂CO₃ (10.65 g; 32.7mmol; 1.5 eq.) and morpholine (2.3 g; 26 mmol; 1.2 eq.) in dioxane (100mL) and the reaction mixture was refluxed for 15 hours. After filtrationthrough a pad of Celite®, the solution was concentrated in vacuo and theresidue purified by column chromatography (petroleum ether/ethylacetate, 80/20) to afford the title compound (4.3 g, 84%) as a yellowsolid.

LC/MS: 236.0 (M+H)⁺. ¹H NMR (CDCl₃, 400 MHz) δ 7.90-7.86 (m, 2H),7.16-7.13 (m, 1H), 4.00-3.98 (m, 4H), 3.91 (s, 3H), 3.12-3.10 (m, 4H),2.45 (s, 3H).

Step 2: 3-Methyl-4-morpholin-4-ylbenzoic acid

Lithium hydroxide (2 g; 84.6 mmol; 5 eq.) was added to a solution ofmethyl 3-methyl-4-morpholin-4-ylbenzoate (4 g; 17 mmol; 1 eq.) in THF(100 mL) and water (5 mL) and the reaction mixture was stirred at 50° C.for 12 hours. The solvent was removed in vacuo, the residue diluted withwater and the aqueous layer washed with DCM (2×50 mL). The aqueous layerwas acidified to pH 4 with conc. HCl and extracted with ethyl acetate(2×100 mL). The combined organic layer was washed with brine, dried oversodium sulfate and concentrated in vacuo to afford the title compound(3.6 g, 97%) as a yellow solid.

HPLC (Method B): Rt 2.36 min (purity 99.3%). LC/MS: 222.1 (M+H)⁺.

¹H NMR (DMSO-d₆, 400 MHz) δ 12.5 (s, 1H), 7.74-7.72 (m, 2H), 7.06-7.04(m, 1H), 3.75-3.72 (m, 4H), 2.90-2.88 (m, 4H), 2.33 (s, 3H).

Intermediate 36 3-cyano-4-(2-methylpiperidin-1-yl)benzoic acid

2-Methylpiperidine (744 mg; 7.5 mmol; 5 eq.) was added to a solution ofmethyl 3-cyano-4-fluorobenzoate (269 mg; 1.5 mmol; 1 eq.) in DMF (2 mL)and the resulting mixture was stirred at room temperature for 2 days.The solution was partitioned between ethyl acetate and water and thephases separated. The organic layer was washed with HCl 0.1 M thenbrine, dried over magnesium sulfate and concentrated in vacuo. Theresidue was taken up in THF (5 mL) and LiOH (126 mg; 3 mmol; 2 eq.) thenwater (5 mL) were added and the reaction mixture was stirred at roomtemperature for 5 hours. The resulting solution was diluted with waterand washed with Et₂O. The aqueous layer was acidified to pH 2 with HCl0.1M and extracted with ethyl acetate. The organic phase was dried overmagnesium sulfate and concentrated in vacuo to afford the title compound(294 mg, 80%) as an off-white solid.

LC/MS: 245.2 (M+H)⁺. ¹H NMR (DMSO-d₆, 300 MHz) δ 12.99 (s, 1H), 8.07 (d,J=2.1 Hz, 1H), 8.00 (dd, J=2.1, 8.8 Hz, 1H), 7.17 (d, J=8.8 Hz, 1H),4.14-4.04 (m, 1H), 3.29-3.21 (m, 2H), 1.87-1.46 (m, 6H), 1.09 (d, J=6.6Hz, 3H)

Intermediate 37 3-methyl-4-(2-methylpyridin-3-yl)benzoic acid

Step 1: methyl3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate

To a suspension of3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid(Synthech BC3558-001, 800 mg; 3.05 mmol; 1 eq.) in MeOH (16 mL) wasadded dropwise thionyl chloride (0.89 mL; 12.2 mmol; 4 eq.) and thereaction mixture was stirred at room temperature for 4 hours. Thesolvent evaporated in vacuo and the crude residue was diluted with Ethylacetate (20 mL). The organic layer was washed with a sat. aq. NaHCO₃ (5mL), water (5 mL) and brine (5 mL), dried over magnesium sulfate andconcentrated in vacuo to afford the title compound (751 mg, 89%) as awhite solid.

HPLC (Method A): Rt 5.28 min (purity 78.4%). LC/MS: 276.9 (M+H)⁺.

Step 2: 3-methyl-4-(2-methylpyridin-3-yl)benzoic acid

A mixture of 3-bromo-2-methylpyridine (0.13 mL; 1.16 mmol; 1 eq.),methyl 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate(353 mg; 1.28 mmol; 1.1 eq.), potassium carbonate (803 mg; 5.81 mmol; 5eq.) and tetrakis(triphenylphosphine)palladium(0) (134 mg; 0.12 mmol;0.1 eq.) in toluene (1 mL) and water (1 mL) was refluxed for 2 hours.The reaction mixture was cooled down to room temperature and filteredthrough a pad of Celite® which was further washed with toluene (20 mL).The filtrate was concentrated in vacuo and the residue taken up in Ethylacetate (10 mL). The organic layer was washed with a sat. aq. NaHCO₃,water and brine, dried over MgSO₄ and concentrated in vacuo. The residue(250 mg; 1.04 mmol; 1 eq.) was taken up in EtOH (7.5 mL) and sodiumhydroxide (5M; 0.62 mL; 3.11 mmol; 3 eq.). The reaction mixture wasstirred at 60° C. for 2 hours then concentrated in vacuo. The residuewas taken up in water (50 mL) and the aqueous phase was washed withEthyl acetate (2×20 mL) then acidified pH 2 with conc. HCl. The solutionwas concentrated in vacuo to ca. 15 mL, the precipitate filtered off andwashed with ACN to afford the title compound (190 mg, 72%) as a beigesolid.

HPLC (Method A): Rt 0.88 min (purity 97.1%). ¹H NMR (DMSO-d6, 300 MHz) δ13.04 (s, 1H), 8.78-8.77 (d, J=5.4 Hz, 1H), 8.16-8.13 (m, 1H), 7.98-7.95(dd, J=1.9, 8.0 Hz, 1H), 7.77-7.76 (m, 2H), 7.56-7.54 (d, J=8.0 Hz, 1H),2.37 (s, 3H), 2.13 (s, 3H).

Intermediate 38 6-(2-methylpiperidin-1-yl)nicotinic acid

-chloronicotinonitrile (Maybridge SPB04745; 1.5 g; 10.8 mmol; 1 eq.) and2-methylpiperidine (25.6 ml; 216.5 mmol; 20 eq.) was stirred at 90° C.for 24 hours. The mixture was diluted with ethyl acetate, washed withwater and brine, dried over magnesium sulphate and concentrated invacuo. To the residue (1.5 g; 7.45 mmol; 1 eq.) in water (90 mL) wasadded potassium hydroxide (2.09 g; 37.3 mmol; 5 eq.) and the reactionmixture was refluxed for 16 hours. After cooling to room temperature,the precipitate was filtered off and the filtrate acidified to pH 6 withHCl 1 M. The precipitate was filtered off and dried to afford the titlecompound as a beige solid.

HPLC (Method A): Rt 3.41 min (purity 97.9%). LC/MS: 221.2 (M+H)⁺.

Intermediate 39 5-methyl-6-(2-methylpiperidin-1-yl)nicotinic acid

Step 1: 5-methyl-6-(2-methylpiperidin-1-yl)nicotinonitrile

A mixture of 5-cyano-2-fluoro-3-methylpyridine (Molekula M52391889; 1.5g; 11 mmol; 1 eq.) and 2-methylpiperidine (5.2 mL; 44.1 mmol; 4 eq.) wasstirred at 90° C. for 16 hours.

The reaction mixture was allowed to return to room temperature thendiluted with ethyl acetate, washed with water, dried over magnesiumsulphate and concentrated in vacuo to afford the title compound (2.2 g,93%) as a brown oil.

HPLC (Method A): Rt 3.60 min (purity 84.5%). LC/MS: 216.2 (M+H)⁺.

Step 2: 5-methyl-6-(2-methylpiperidin-1-yl)nicotinic acid

A mixture of 5-methyl-6-(2-methylpiperidin-1-yl)nicotinonitrile (1 g;4.64 mmol; 1 eq.) and potassium hydroxide (1.3 g; 23.2 mmol; 5 eq.) inwater (60 mL) was stirred at reflux for 16 hours. The pH was adjusted to5-6 with HCl 1 M and the aqueous layer extracted with ethyl acetate. Thecombined organic phase was dried over magnesium sulphate andconcentrated in vacuo to afford the title compound (1.1 g, 100%) as ayellow oil.

HPLC (Method A): Rt 1.82 min (purity 88.7%). LC/MS: 235.2 (M+H)⁺.

Intermediate 40 6-[2-(methoxymethyl)pyrrolidin-1-yl]-5-methylnicotinicacid

[2-(methoxymethyl)pyrrolidin-1-yl]-5-methylnicotinonitrile

2-(Methoxymethyl)pyrrolidine (Acb-blocks C5A-0029; 406 mg; 3.53 mmol;1.2 eq.) and DIEA (1.52 ml; 8.82 mmol; 3 eq.) were added to a solutionof 5-cyano-2-fluoro-3-methylpyridine (400 mg; 2.94 mmol; 1 eq.) in1-butanol (1 mL) and the reaction mixture was stirred at 90° C. for 16hours. The reaction mixture was allowed to return to room temperaturethen diluted with ethyl acetate, washed with water, dried over magnesiumsulphate and concentrated in vacuo to afford the title compound (0.59 g,87%) as a yellow oil.

HPLC (Method A): Rt 2.42 min (purity 99.1%). LC/MS: 200.1 (M+H)⁺.

Step 2: 6-[2-(methoxymethyl)pyrrolidin-1-yl]-5-methylnicotinic acid

A mixture of6-[2-(methoxymethyl)pyrrolidin-1-yl]-5-methylnicotinonitrile (591 mg;2.56 mmol; 1 eq.) and potassium hydroxide (717 mg; 12.8 mmol; 5 eq.) inwater (20 mL) was stirred at reflux for 16 hours. The pH was adjusted to5-6 with HCl 1M and the aqueous layer extracted with ethyl acetate. Thecombined organic phase was dried over magnesium sulphate andconcentrated in vacuo to afford the title compound (0.63 g, 99%) as awhite solid.

HPLC (Method A): Rt 1.44 min (purity 96.6%). LC/MS: 219.1 (M+H)⁺.

Intermediate 41 2-(hydroxymethyl)-2′-methylbiphenyl-4-carboxylic acid

Step 1: Methyl 4-bromo-3-(bromomethyl)benzoate

To a solution of methyl 4-bromo-3-methylbenzoate (Aldrich 532878, 50 g;218 mmol; 1 eq.) in CHCl₃ (1 L) were added NBS (46.6 g; 262 mmol; 1.2eq.) in one portion and α,α′-azoisobutyronitrile (0.72 g; 4.37 mmol;0.02 eq.) and the reaction mixture was stirred at 70° C. for 2 days. Themixture was cooled down to room temperature and water (500 mL) wasadded. The organic layer was washed with aq. NaHCO₃ (400 mL), then brine(500 mL), dried over MgSO₄ and concentrated in vacuo. The residue waswashed with n-pentane (2×500 mL) to afford the title compound as ayellow solid.

HPLC (Method A): Rt 4.44 min (purity 97.9%). ¹H NMR (DMSO-d₆, 400 MHz) δ8.24 (d, J=1.9 Hz, 1H), 7.88-7.82 (m, 2H), 4.87 (s, 2H), 3.91 (s, 3H).

Step 2: Methyl 3-[(acetyloxy)methyl]-4-bromobenzoate

To a solution of methyl 4-bromo-3-(bromomethyl)benzoate (6.5 g; 21 mmol;1 eq.) in AcOH (32.5 mL) was added sodium acetate (3.46 g; 42 mmol; 2eq.) and the reaction mixture was stirred at 100° C. for 12 hours. Afterconcentration in vacuo, the residue was partitioned between Ethylacetate and water. The organic layer was washed with 5% aq. NaHCO₃ thenbrine, dried over MgSO₄ and concentrated in vacuo. Purification bycolumn chromatography c-hexane/ethyl acetate, 5/1) afforded the titlecompound (4.78 g, 79%) as a white solid

HPLC (Method A): Rt 4.37 min (purity 98.1%). ¹H NMR (DMSO, 400 MHz) δ8.03 (m, 1H), 7.85-7.84 (d, J=1.3 Hz, 1H), 5.18 (s, 2H), 3.87 (s, 3H),2.11 (s, 3H).

Step 3: Methyl 2-[(acetyloxy)methyl]-2′-methylbiphenyl-4-carboxylate

A mixture of methyl 3-[(acetyloxy)methyl]-4-bromobenzoate (4.7 g; 16.4mmol; 1 eq.), o-tolylboronic acid (2.45 g; 18 mmol; 1.1 eq.), potassiumcarbonate (11.3 g; 82 mmol; 5 eq.) and Pd(PPh₃)₄ (1.89 g; 1.64 mmol; 0.1eq.) in toluene (23.5 mL) and water (23.5 mL) was refluxed for 2 hours.After cooling to room temperature, the reaction mixture was filteredthrough a pad of Celite® which was further washed with toluene (50 mL).The filtrate was concentrated in vacuo, the residue taken up in ethylacetate (250 mL) and washed with sat. aq. NaHCO₃ (100 mL), water (100mL) and brine (100 mL), dried over magnesium sulphate and concentratedin vacuo to afford the title compound (4.9 g, 100%) as a brown oil.

HPLC (Method A): Rt 5.23 min (purity 62.3%).

Step 4: 2-(hydroxymethyl)-2′-methylbiphenyl-4-carboxylic acid

Sodium hydroxide (5M; 12.1 mL; 60.3 mmol; 3 eq.) was added to a solutionof methyl 2-[(acetyloxy)methyl]-2′-methylbiphenyl-4-carboxylate (6 g; 20mmol; 1 eq.) in EtOH (180 mL) and the reaction mixture was stirred at60° C. for 2 hours. After concentration in vacuo, the residue was takenup in water (500 mL) and washed with Ethyl acetate (2×100 mL). Theaqueous phase was acidified to pH 2 with conc. HCl and extracted withEthyl acetate (2×100 mL). The combined organic layer was dried overMgSO₄ and concentrated in vacuo to afford the title compound (3.46 g,71%) as a yellow solid.

HPLC (Method A): Rt 3.77 min (purity 96.1%). LC/MS: 241.2 (M−H)⁻. ¹H NMR(DMSO-d₆, 400 MHz) δ 12.97 (br s, 1H), 8.20-8.19 (m, 1H), 7.87-7.84 (dd,J=8.0 Hz, 1.86 Hz, 1H), 7.37-7.06 (m, 5H), 5.23-5.19 (m, 1H), 4.25-4.09(m, 2H), 2.01 (s, 3H

Intermediate 42 4-Piperidin-1-yl-3-(trifluoromethyl)benzoic acid

Step 1: 4-Piperidin-1-yl-3-(trifluoromethyl)benzonitrile

A mixture of 4-fluoro-3-trifluoro-methylbenzonitrile (5 g; 26.4 mmol; 1eq.) and piperidine (5.2 mL; 52.8 mmol; 2 eq.) was stirred at 100° C. 20hours. The reaction mixture was cooled, diluted with water (200 mL) andextracted with ethyl acetate (2×200 mL). The combined organic layer waswashed with water (100 mL) and brine (100 mL), dried over sodiumsulphate and concentrated in vacuo. The residue was purified by columnchromatography (petroleum ether/ethyl acetate, 85/15) to afford thetitle compound of the title compound as an off-white solid.

HPLC (Method B): Rt 4.80 min (purity 99.1%).

LC/MS: 255.1 (M+H)⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 7.87 (s, 1H), 7.72-7.70(m, 1H), 7.27-7.24 (m, 1H), 3.02-2.97 (m, 4H), 1.85-1.80 (m, 4H),1.75-1.70 (m, 2H).

Step 2: Methyl 4-piperidin-1-yl-3-(trifluoromethyl)benzoate

A mixture of 4-piperidin-1-yl-3-(trifluoromethyl)benzonitrile (7.4 g, 27mmol) and HCl in methanol (4M, 250 mL) was stirredat 60° C. for 12hours, then concentrated to dryness. The residue was partitioned betweenethyl acetate and 10% aq. NaHCO₃, and the organic layer washed withwater then brine. Drying over magnesium sulphate and concentration invacuo afforded the title compound (6.6 g, 85%) as a yellow oil.

¹H NMR (DMSO-d₆, 400 MHz) δ 8.38 (s, 1H), 8.25-8.21 (m, 1H), 7.47-7.45(m, 1H), 3.94 (s, 3H), 3.04-2.99 (m, 4H), 1.81-1.78 (m, 4H), 1.75-1.70(m, 2H).

Step 3: 4-Piperidin-1-yl-3-(trifluoromethyl)benzoic acid

Lithium hydroxide (2.83 g, 67.5 mmol, 2 eq.) was added to a solution ofmethyl 4-piperidin-1-yl-3-(trifluoromethyl)benzoate

(9.7 g, 33.7 mmol, 1 eq.) in THF (50 mL) and water (5 mL) and thereaction mixture was stirred at room temperature for 12 hours. Thesolvent was removed in vacuo and the residue taken up in water. Theaqueous layer was washed with DCM, acidified to pH 2 with conc. HCl andextracted with ethyl acetate. The organic phase was washed with brine,dried over sodium sulphate and concentrated in vacuo to afford the titlecompound (3.0 g, 95%) as a white solid. HPLC (Method B): Rt 4.36 min(purity 91.7%). LC/MS: 272.9 (M+H)⁺. ¹H NMR (DMSO-d₆, 400 MHz) δ 13.30(s, 1H), 8.12-8.10 (m, 2H), 7.49-7.47 (m, 1H), 2.92-2.90 (m, 4H),1.65-1.60 (m, 4H), 1.57-1.53 (m, 2H).

Example 11-{4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}piperidine

Trichloroacetonitrile (0.08 mL; 0.75 mmol; 1.50 eq.), intermediate 4(125 mg; 0.50 mmol; 1 eq.), polymer bound triphenylphosphine (499.49 mg;1.50 mmol; 3 eq.), intermediate 1 (91.31 mg; 0.55 mmol; 1.10 eq.) andDIEA (0.17 mL; 1 mmol; 2 eq.) were reacted according to generalprocedure 1. Purification by column chromatography (c-hexane/ethylacetate, 80/20) followed by crystallization (Et₂O/n-hexane) afforded thetitle compound as an orange solid.

HPLC (Method A): Rt 5.32 min (purity 97.4%). LC/MS: 381.0 (M+H)⁺.

Example 24-{4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}morpholine

Trichloroacetonitrile (0.08 mL; 0.75 mmol; 1.50 eq.), intermediate 5(126 mg; 0.50 mmol; 1 eq.), polymer bound triphenylphosphine (499.55 mg;1.50 mmol; 3 eq.), intermediate 1 (91.32 mg; 0.55 mmol; 1.10 eq.) andDIEA (0.17 mL; 1 mmol; 2 eq.) were reacted according to generalprocedure 1. Purification by column chromatography using DCM as eluentafforded an orange oil which crystallized on standing to afford thetitle compound as a yellow solid.

HPLC (Method A): Rt 4.35 min (purity 93.4%). LC/MS: 383.0 (M+H)⁺.

Example 31-(4-{3-[2-(Trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)piperidine

Oxalyl chloride (166.96 mg; 1.32 mmol; 3 eq.), intermediate 6 (90 mg;0.44 mmol; 1 eq.), intermediate 2 (96.53 mg; 0.44 mmol; 1 eq.) and DIEA(170.01 mg; 1.32 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 90/10) afforded the title compound as an off-white solid.

HPLC (Method A): Rt 5.39 min (purity 99.5%). LC/MS: 390.0 (M+H)⁺.

Example 41-{4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]phenyl}piperidine

Oxalyl chloride (166.96 mg; 1.32 mmol; 3 eq.), intermediate 6 (90 mg;0.44 mmol; 1 eq.), intermediate 1 (72.87 mg; 0.44 mmol; 1 eq.) and DIEA(170.01 mg; 1.32 mmol; 3 eq.) in THF (1 mL) were reacted according togeneral procedure 2. Purification by crystallization (Et₂O/n-hexane)afforded the title compound as an off-white solid.

HPLC (Method A): Rt 4.10 min (purity 95.0%). LC/MS: 336.0 (M+H)⁺.

Example 5N-Cyclohexyl-N-methyl-2-nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 7 (139.15mg; 0.50 mmol; 1 eq), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography α-hexane/ethylacetate, 90/10) followed by crystallization (n-hexane) afforded thetitle compound as a yellow solid.

HPLC (Method A): Rt 6.44 min (purity 98.6%). LC/MS: 463.2 (M+H)⁺.

Example 65-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-morpholin-4-ylaniline

Stannous chloride dihydrate (147.53 mg; 0.65 mmol; 5 eq.) was added to asolution of example 2 (50 mg; 0.13 mmol; 1 eq.) in EtOH (10 mL), and thereaction mixture was stirred at 70° C. for 3 hours. After cooling, thesolution was partitioned between ethyl acetate and aqueous NaHCO₃. Theorganic layer was washed four times with brine, dried over magnesiumsulfate and concentrated in vacuo to give a yellow solid. The latter wastriturated in Et₂O and filtrated to afford the title compound (38 mg,82%) as a pale yellow solid.

HPLC (Method A): Rt 3.60 min (purity 96.7%). LC/MS: 353.1 (M+H)⁺.

Example 71-{4-[3-(2-Ethoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}piperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 4 (125.13mg; 0.50 mmol; 1 eq.), intermediate 3 (90.10 mg; 0.50 mmol; 1 eq.) andDIEA (0.17 mL; 1 mmol; 2 eq.) were reacted according to generalprocedure 2. Purification by chromatography (c-hexane/ethyl acetate,95/5 then 90/10) afforded the title compound (139 mg, 70%) as an orangeoil.

HPLC (Method A): Rt 5.73 min (purity 98.0%). LC/MS: 395.1 (M+H)⁺.

Example 82-Methyl-1-(2-nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)piperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 8 (132.14mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-Hexane/ethylacetate, 90/10) followed by crystallization (Et₂O/n-hexane) afforded thetitle compound as a yellow solid.

HPLC (Method A): Rt 6.26 min (purity 98.4%). LC/MS: 449.2 (M+H)⁺.

Example 91-{4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}-2-methylpiperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 8 (132.14mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound (162 mg, 82%) as an orangeoil.

HPLC (Method A): Rt 5.46 min (purity 98.0%). LC/MS: 395.1 (M+H)⁺.

Example 101-{4-[3-(2-Ethoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}-2-methylpiperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 8 (132.14mg; 0.50 mmol; 1 eq.), intermediate 3 (90.10 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 90/10) afforded the title compound as an orange oil.

HPLC (Method A): Rt 5.98 min (purity 97.6%). LC/MS: 409.2 (M+H)⁺.

Example 113,3-Difluoro-1-(2-nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)piperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 9 (143.12mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (ethyl acetate/n-hexane)afforded the title compound (218 mg, 93%) as a yellow solid.

HPLC (Method A): Rt 5.64 min (purity 99.1%). LC/MS: 470.9 (M+H)⁺.

Example 123,3-Difluoro-1-{4-[3-(2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}piperidine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 9 (143.12mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (ethyl acetate/n-hexane)afforded the title compound as a yellow solid.

HPLC (Method A): Rt 4.97 min (purity 99.4%). LC/MS: 417.0 (M+H)⁺.

Example 133-(2-Methoxyphenyl)-5-(3-nitro-4-pyrrolidin-1-ylphenyl)-1,2,4-oxadiazole

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 10 (118.11mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound as an orange solid.

HPLC (Method A): Rt 4.82 min (purity 96.7%). LC/MS: 366.9 (M+H)⁺.

Example 145-(3-Nitro-4-pyrrolidin-1-ylphenyl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 10 (118.11mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound as an orange solid.

HPLC (Method A): Rt 5.64 min (purity 93.1%). LC/MS: 420.9 (M+H)⁺.

Example 151-(2-Nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)azepane

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 11 (132.14mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound as a yellow solid.

HPLC (Method A): Rt 6.07 min (purity 87.8%). LC/MS: 448.9 (M+H)⁺.

Example 161-{4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}azepane

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 11 (132.14mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound as an orange solid.

HPLC (Method A): Rt 5.29 min (purity 88.5%). LC/MS: 395.0 (M+H)⁺.

Example 174-(2-Nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)morpholine

Oxalyl chloride (131 μl; 1.55 mmol; 3 eq.), intermediate 5 (130 mg; 0.52mmol; 1 eq.), intermediate 2 (113.47 mg; 0.52 mmol; 1 eq.) and DIEA(114.20 μl; 1.55 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 80/20) afforded the title compound as a yellow solid.

HPLC (Method A): Rt 5.24 min (purity 96.7%). LC/MS: 436.9 (M+H)⁺.

Example 184-[4-[3-(2-Methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(trifluoromethyl)phenyl]morpholine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 14 (137.61mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (ethyl acetate/n-pentane)afforded the title compound (160 mg, 79%) as an off-white solid. HPLC(Method A): Rt 4.98 min (purity 95.7%). LC/MS: 406.1 (M+H)⁺.

Example 194-[4-{3-[2-(Trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}-2-(trifluoromethyl)phenyl]morpholine

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 14 (137.61mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (n-pentane) afforded thetitle compound as an off-white solid.

HPLC (Method A): Rt 5.78 min (purity 98.9%). LC/MS: 459.9 (M+H)⁺.

Example 205-(2,2′-Dimethylbiphenyl-4-yl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Trichloroacetonitrile (0.08 mL; 0.75 mmol; 1.50 eq.), intermediate 16(113 mg; 0.50 mmol; 1 eq.), polymer bound triphenylphosphine (499.40 mg;1.50 mmol; 3 eq.), intermediate 2 (91.29 mg; 0.55 mmol; 1.10 eq.) andDIEA (0.17 mL; 1 mmol; 2 eq.) in THF (2 mL) were reacted according togeneral procedure 1. Purification by column chromatography(c-hexane/ethyl acetate, 90/10) afforded the title compound as abrownish oil.

HPLC (Method A): Rt 5.87 min (purity 89.7%). LC/MS: 357.0 (M+H)⁺.

Example 215-(2,2′-Dimethylbiphenyl-4-yl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 16 (113.14mg; 0.50 mmol; 1 eq), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane thenc-hexane/ethyl acetate, 98/2) afforded the title compound as acolourless oil.

HPLC (Method A): Rt 6.62 min (purity 99.1%). LC/MS: 411.1 (M+H)⁺.

Example 225-(2,2′-Dimethylbiphenyl-4-yl)-3-(2-ethoxyphenyl)-1,2,4-oxadiazole

Oxalyl chloride (0.04 mL; 0.52 mmol; 1.05 eq.), intermediate 16 (113.14mg; 0.50 mmol; 1 eq), intermediate 3 (90.10 mg; 0.50 mmol; 1 eq.) andDIEA (0.17 mL; 1 mmol; 2 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 97/3 to 95/5) followed by crystallization (ethanol/methanol)afforded the title compound as a white solid.

HPLC (Method A): Rt 6.29 min (purity 99.7%). LC/MS: 371.1 (M+H)⁺.

Example 235-[2-Methyl-2′-(trifluoromethyl)biphenyl-4-yl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 18 (140.12mg; 0.50 mmol; 1 eq.), intermediate 2 (110.08 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (n-hexane) the titlecompound as a white solid.

HPLC (Method A): Rt 6.45 min (purity 99.6%). LC/MS: 465.0 (M+H)⁺.

Example 243-(2-Methoxyphenyl)-5-[2-methyl-2′-(trifluoromethyl)biphenyl-4-yl]-1,2,4-oxadiazole

Oxalyl chloride (190.39 mg; 1.50 mmol; 3 eq.), intermediate 18 (140.12mg; 0.50 mmol; 1 eq.), intermediate 1 (83.09 mg; 0.50 mmol; 1 eq.) andDIEA (193.87 mg; 1.50 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 90/10) afforded the title compound (152 mg, 74%) as anoff-white solid.

HPLC (Method A): Rt 5.92 min (purity 95.2%). LC/MS: 411.4 (M+H)⁺.

Example 253-(2-Methoxyphenyl)-5-[3-methyl-4-(4-methyl-3-thienyl)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (131.13 μl; 1.55 mmol; 3 eq.), intermediate 20 (120 mg;0.52 mmol; 1 eq.), intermediate 1 (85.84 mg; 0.52 mmol; 1 eq.) and DIEA(267.05 μl; 1.55 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by column chromatography (c-hexane/ethylacetate, 90/10) followed by crystallization (n-pentane) afforded thetitle compound as a white solid.

HPLC (Method A): Rt 5.68 min (purity 99.9%). LC/MS: 362.9 (M+H)⁺.

Example 263-(2-Methoxyphenyl)-5-(2′-methyl-2-nitrobiphenyl-4-yl)-1,2,4-oxadiazole

Oxalyl chloride (118.42 μl; 1.40 mmol; 3 eq.), intermediate 22 (120 mg;0.47 mmol; 1 eq.), intermediate 1 (77.52 mg; 0.47 mmol; 1 eq.) and DIEA(103.36 μl; 1.40 mmol; 3 eq.) were reacted according to generalprocedure 2. Purification by crystallization (n-pentane) afforded thetitle compound as a white solid.

HPLC (Method A): Rt 5.37 min (purity 94.6%). LC/MS: 387.9 (M+H)⁺.

Example 27 5-(4-Cyclohexylphenyl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Trichloroacetonitrile (0.15 ml; 1.50 mmol; 1.50 eq.),4-cyclohexylbenzoic acid (Lancaster 5086; 204.27 mg; 1.00 mmol; 1 eq.),polymer bound triphenylphosphine (1.87 g; 3.00 mmol; 3 eq.),intermediate 1 (182.80 mg; 1.10 mmol; 1.10 eq.) and DIEA (0.34 ml; 2.00mmol; 2 eq.) were reacted according to general procedure 1. Purificationby column chromatography (c-hexane/ethyl acetate, 98/2 to 95/5) followedby crystallization (methanol) afforded the title compound as a yellowsolid.

HPLC (Method A): Rt 5.98 min (purity 99.9%). LC/MS: 335.1 (M+H)⁺.

Example 28 5-(4-Isobutylphenyl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Trichloroacetonitrile (0.15 mL; 1.50 mmol; 1.50 eq.), 4-isobutylbenzoicacid (Enamine EN300-11610; 204.27 mg; 1.00 mmol; 1 eq.), polymer boundtriphenylphosphine (1.87 g; 3.00 mmol; 3 eq.), intermediate 1 (182.80mg; 1.10 mmol; 1.10 eq.) and DIEA (0.34 mL; 2.00 mmol; 2 eq.) werereacted according to general procedure 1. Purification bycrystallization (CH₃CN/water) afforded the title compound as a whitesolid.

HPLC (Method A): Rt 5.67 min (purity 100%). LC/MS: 309.0 (M+H)⁺.

Example 295-(4-Cyclohexylphenyl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (0.04 mL; 0.52 mmol; 1.05 eq.), 4-cyclohexylbenzoic acid(Lancaster 5086; 89.11 mg; 0.50 mmol; 1.00 eq.), intermediate 2 (110.08mg; 0.50 mmol; 1 eq.) and DIEA (0.17 mL; 1.40 mmol; 3 eq.) were reactedaccording to general procedure 2. Purification by column chromatography(c-hexane/ethyl acetate, 80/20) afforded the title compound as a whitesolid.

HPLC (Method A): Rt 6.66 min (purity 96.8%). LC/MS: 389.1 (M+H)⁺.

Example 305-(4-Isobutylphenyl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (0.04 mL; 0.52 mmol; 1.05 eq.), 4-isobutylbenzoic acid(Enamine EN300-11610; 102.13 mg; 0.50 mmol; 1.00 eq.), intermediate 2(110.08 mg; 0.50 mmol; 1 eq.) and DIEA (0.17 mL; 1.40 mmol; 3 eq.) werereacted according to general procedure 2. Purification by columnchromatography (c-hexane/ethyl acetate, 99/1) afforded the titlecompound as a colorless oil.

HPLC (Method A): Rt 6.39 min (purity 99.1%). LC/MS: 363.0 (M+H)⁺.

Example 311-(2-nitro-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)piperidine

Oxalyl chloride (71 μL; 0.84 mmol; 3 eq.), Intermediate 4 (70 mg; 0.28mmol; 1 eq.), Intermediate 2 (62 mg; 0.28 mmol, 1 eq.) and DIEA (145 μL;0.84 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 80/20)followed by precipitation in n-pentane afforded the title compound as ayellow solid.

HPLC (Method A): Rt 6.11 min (purity 100%). LC/MS: 434.9 (M+H)⁺.

Example 321-{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitrophenyl}-2-methylpiperidine

Intermediate 23 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg; 1.5 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 80/20) followed bycrystallisation from ethyl acetate/n-pentane afforded the title compoundas a yellow solid.

HPLC (Method A): Rt 5.64 min (purity 99.2%). LC/MS: 412.9 (M+H)⁺.

Example 335-(2,2′-dimethylbiphenyl-4-yl)-3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazole

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 16 (113 mg; 0.5mmol; 1 eq.), Intermediate 23 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 90/10)afforded the title compound as a yellow solid.

HPLC (Method A): Rt 5.96 min (purity 100%). LC/MS: 453.9 (M+H)⁺.

Example 342-methyl-1-[4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}-2-(trifluoromethyl)phenyl]piperidine

Oxalyl chloride (106 μL; 1.25 mmol; 3 eq.), Intermediate 24 (120 mg;0.42 mmol; 1 eq.), Intermediate 2 (92 mg; 0.42 mmol, 1 eq.) and DIEA (93μL; 1.25 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 80/20)followed by precipitation from n-pentane afforded the title compound asa yellow solid.

HPLC (Method A): Rt 7.09 min (purity 93.5%). LC/MS: 471.8 (M+H)⁺.

Example 351-[4-[3-(2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(trifluoromethyl)phenyl]-2-methylpiperidine

Oxalyl chloride (106 μL; 1.25 mmol; 3 eq.), Intermediate 24 (120 mg;0.42 mmol; 1 eq.), Intermediate 1 (69 mg; 0.42 mmol, 1 eq.) and DIEA (93μL; 1.25 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 80/20)followed by precipitation from n-pentane afforded the title compound asa white solid.

HPLC (Method A): Rt 6.47 min (purity 99.2%). LC/MS: 417.6 (M+H)⁺.

Example 365-(2′-methyl-2-nitrobiphenyl-4-yl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (118 μL; 1.40 mmol; 3 eq.), Intermediate 22 (120 mg;0.47 mmol; 1 eq.), Intermediate 2 (103 mg; 0.47 mmol, 1 eq.) and DIEA(103 μL; 1.40 mmol; 3 eq.) were reacted according to general procedure2. Purification by column chromatography (c-hexane/ethyl acetate, 90/10)afforded the title compound as a yellow oil.

HPLC (Method A): Rt 6.13 min (purity 98.3%). LC/MS: 441.9 (M+H)⁺.

Example 37N-dimethanesulfonyl-2-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(methoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline

Step 1:2-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(methoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline

Stannous chloride dihydrate (285 mg; 1.26 mmol; 5 eq.) was added to asolution of Example 12 (105 mg; 0.25 mmol; 1 eq.) in EtOH (20 mL) andthe resulting mixture was stirred at 70° C. for 3 hours, then at roomtemperature for 16 hours. The solution was diluted with sat. aq. NaHCO₃and extracted with ethyl acetate. The combined organic layer was washedwith brine, dried over magnesium sulfate and concentrated in vacuo toafford the title compound (97 mg, 100%) as a yellow solid.

HPLC (Method A): Rt 4.61 min (purity 93.3%). LC/MS: 387.2 (M+H)⁺.

Step 2:N-dimethanesulfonyl-2-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(methoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline

Methanesulfonyl chloride (32 mg; 0.28 mmol; 1.2 eq.) was added dropwiseto a suspension of2-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline(90 mg; 0.23 mmol; 1 eq.) and DIEA (60 mg; 0.47 mmol; 2 eq.) in DCM (5mL) and the resulting mixture was stirred at room temperature for 3days. After evaporation of the solvent, the residue was purified byflash chromatography (c-hexane/ethyl acetate, 70/30) followedcrystallization from Et₂O to afford the title compound as an off-whitesolid.

HPLC (Method A): Rt 4.73 min (purity 100%). LC/MS: 542.9 (M+H)⁺.

Example 385-[4-(2,5-dimethylpyrrolidin-1-yl)-3-nitrophenyl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (152 μL; 1.2 mmol; 3 eq.), Intermediate 25 (106 mg; 0.4mmol; 1 eq.), Intermediate 2 (88 mg; 0.4 mmol, 1 eq.) and DIEA (155 μL;1.2 mmol; 3 eq.) were reacted according to general procedure 2.Purification by crystallisation from ethyl acetate/n-pentane affordedthe title compound as a yellow solid.

HPLC (Method A): Rt 6.15 min (purity 100%). LC/MS: 449.1 (M+H)⁺.

Example 395-[4-(2,5-dimethylpyrrolidin-1-yl)-3-nitrophenyl]-3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazole

Intermediate 23 (74 mg; 0.4 mmol, 1 eq.) and DIEA (155 μL; 1.2 mmol; 3eq.) were reacted according to general procedure 2. Purification bycrystallisation from ethyl acetate/n-pentane afforded the title compoundas a yellow solid.

HPLC (Method A): Rt 5.54 min (purity 99.3%). LC/MS: 413.1 (M+H)⁺.

Example 405-(2′-methoxy-2-methylbiphenyl-4-yl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Intermediate 1 (82 mg; 0.5 mmol, 1 eq.) and DIEA (256 μL; 1.5 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 90/10) followed byprecipitation from n-pentane afforded the title compound as a whitesolid.

HPLC (Method A): Rt 5.51 min (purity 97.7%). LC/MS: 373.0 (M+H)⁺.

Example 412-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}aniline

Stannous chloride dihydrate (478 mg; 2.13 mmol; 5 eq.) was added to asolution of Example 11 (200 mg; 0.43 mmol; 1 eq.) in EtOH (20 mL) andthe resulting mixture was stirred at 70° C. for 3 hours, then at roomtemperature for 16 hours. The solution was diluted with sat. aq. NaHCO₃and extracted with ethyl acetate. The combined organic layer was washedwith brine, dried over magnesium sulfate and concentrated in vacuo.Purification by column chromatography (c-hexane/ethyl acetate, 70/30)followed by crystallization from n-pentane afforded the title compoundas an off-white solid.

HPLC (Method A): Rt 5.54 min (purity 99.5%). LC/MS: 440.9 (M+H)⁺.

Example 425-[4-(2,5-dimethylpyrrolidin-1-yl)-3-nitrophenyl]-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Intermediate 1 (66 mg; 0.4 mmol, 1 eq.) and DIEA (155 mg; 1.2 mmol; 3eq.) were reacted according to general procedure 2. Purification byprecipitation from Et₂O/n-pentane afforded the title compound as ayellow solid.

HPLC (Method A): Rt 5.38 min (purity 99.1%). LC/MS: 394.9 (M+H)⁺.

Example 43N-(2-(3,3-difluoropiperidin-1-yl)-5-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)methanesulfonamide

Methanesulfonylchloride (29.23 mg; 0.26 mmol; 0.6 eq.) was added to asolution of Example 41 (75 mg; 0.43 mmol; 1 eq.) in pyridine (1 mL) andthe resulting solution was stirred at room temperature for 36 hours thenconcentrated in vacuo. The residue was purified by column chromatography(c-hexane/ethyl acetate, 70/30) followed by crystallization from ethylacetate/n-pentane to afford the title compound as an off-white solid.

HPLC (Method A): Rt 5.31 min (purity 100%). LC/MS: 518.6 (M+H)⁺.

Example 445-[3-methyl-4-(4-methyl-3-thienyl)phenyl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

oride (131 μL; 1.55 mmol; 3 eq.), Intermediate 20 (120 mg; 0.52 mmol; 1eq.), Intermediate 2 (114 mg; 0.52 mmol, 1 eq.) and DIEA (267 μL; 1.55mmol; 3 eq.) were reacted according to general procedure 2. Purificationby column chromatography (c-hexane/ethyl acetate, 95/5) afforded thetitle compound as a white solid.

HPLC (Method A): Rt 6.57 min (purity 91.5%). LC/MS: 417.0 (M+H)⁺.

Example 455-(2′-methoxy-2-methylbiphenyl-4-yl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (126 μL; 1.49 mmol; 3 eq.), Intermediate 26 (120 mg; 0.5mmol; 1 eq.), Intermediate 2 (109 mg; 0.5 mmol, 1 eq.) and DIEA (256 μL;1.49 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 95/5)afforded the title compound as a yellow oil.

HPLC (Method A): Rt 6.16 min (purity 90.6%). LC/MS: 427.0 (M+H)⁺.

Example 465-(2-methoxy-2′-methylbiphenyl-4-yl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

μL; 1.49 mmol; 3 eq.), Intermediate 27 (120 mg; 0.5 mmol; 1 eq.),Intermediate 1 (82 mg; 0.5 mmol, 1 eq.) and DIEA (256 μL; 1.49 mmol; 3eq.) were reacted according to general procedure 2. Purification byprecipitation from DCM/n-pentane afforded the title compound as a brownsolid.

HPLC (Method A): Rt 5.46 min (purity 95.9%). LC/MS: 372.9 (M+H)⁺.

Example 475-(2′,4′-dimethoxy-2-methylbiphenyl-4-yl)-3-(2-methoxyphenyl)-1,2,4-oxadiazole

Oxalyl chloride (112 μL; 1.32 mmol; 3 eq.), Intermediate 28 (120 mg;0.44 mmol; 1 eq.), Intermediate 1 (73 mg; 0.44 mmol, 1 eq.) and DIEA(228 μL; 1.32 mmol; 3 eq.) were reacted according to general procedure2. Purification by precipitation from DCM/n-pentane afforded the titlecompound as a yellow solid.

HPLC (Method A): Rt 5.53 min (purity 98.1%). LC/MS: 403.1 (M+H)⁺.

Example 485-[3-methoxy-4-(4-methyl-3-thienyl)phenyl]-3-(2-methoxyphenyl)-1,2,4-oxadiazole

μL; 1.45 mmol; 3 eq.), Intermediate 29 (120 mg; 0.48 mmol; 1 eq.),Intermediate 1 (80 mg; 0.48 mmol, 1 eq.) and DIEA (250 μL; 1.45 mmol; 3eq.) were reacted according to general procedure 2. Purification byprecipitation from DCM/n-pentane afforded the title compound as a whitesolid.

HPLC (Method A): Rt 5.49 min (purity 96.4%). LC/MS: 378.9 (M+H)⁺.

Example 495-[4-(3,5-dimethylisoxazol-4-yl)-3-methylphenyl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (132 μL; 1.56 mmol; 3 eq.), Intermediate 30 (120 mg;0.52 mmol; 1 eq.), Intermediate 2 (114 mg; 0.52 mmol, 1 eq.) and DIEA(268 μL; 1.56 mmol; 3 eq.) were reacted according to general procedure2. Purification by column chromatography (c-hexane/ethyl acetate, 80/20)afforded the title compound as a yellow oil.

HPLC (Method A): Rt 5.71 min (purity 96.7%). LC/MS: 415.9 (M+H)⁺.

Example 505-[4-(3,5-dimethylisoxazol-4-yl)-3-methylphenyl]-3-(2-methoxyphenyl)-1,2,4-oxadiazole

μL; 1.56 mmol; 3 eq.), Intermediate 30 (120 mg; 0.52 mmol; 1 eq.),Intermediate 1 (86 mg; 0.52 mmol, 1 eq.) and DIEA (268 μL; 1.56 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 80/20) afforded the titlecompound as an orange foam.

HPLC (Method A): Rt 4.85 min (purity 93.8%). LC/MS: 361.9 (M+H)⁺.

Example 514-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-N,N-bis(2-methoxyethyl)-2-nitroaniline

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 31 (149 mg; 0.5mmol; 1 eq.), Intermediate 23 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 50/50)followed by crystallisation from Et₂O/n-pentane afforded the titlecompound as a yellow solid.

HPLC (Method A): Rt 4.72 min (purity 96.6%). LC/MS: 447.1 (M+H)⁺.

Example 52N,N-bis(2-methoxyethyl)-4-[3-(2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-nitroaniline

Intermediate 1 (83 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg; 1.5 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 50/50) followed bycrystallisation from Et2O/n-pentane afforded the title compound as ayellow solid.

HPLC (Method A): Rt 4.55 min (purity 98.0%). LC/MS: 429.1 (M+H)⁺.

Example 535-(2-methoxy-2′-methylbiphenyl-4-yl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

μL; 1.49 mmol; 3 eq.), Intermediate 27 (120 mg; 0.5 mmol; 1 eq.),Intermediate 2 (109 mg; 0.5 mmol, 1 eq.) and DIEA (256 μL; 1.5 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 95/5) afforded the titlecompound as a white solid.

HPLC (Method A): Rt 6.39 min (purity 98.0%). LC/MS: 427.0 (M+H)⁺.

Example 545-[3-methoxy-4-(4-methyl-3-thienyl)phenyl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

μL; 1.45 mmol; 3 eq.), Intermediate 29 (120 mg; 0.48 mmol; 1 eq.),Intermediate 2 (106 mg; 0.48 mmol, 1 eq.) and DIEA (250 μL; 1.45 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 95/5) afforded the titlecompound as a yellow oil.

HPLC (Method A): Rt 6.15 min (purity 98.1%). LC/MS: 432.8 (M+H)⁺.

Example 555-[2′-methyl-2-(trifluoromethyl)biphenyl-4-yl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (109 μL; 1.28 mmol; 3 eq.), Intermediate 34 (120 mg;0.43 mmol; 1 eq.), Intermediate 2 (94 mg; 0.43 mmol, 1 eq.) and DIEA(221 μL; 1.28 mmol; 3 eq.) were reacted according to general procedure2. Purification by column chromatography (c-hexane/ethyl acetate, 95/5)afforded the title compound as a white solid.

HPLC (Method A): Rt 6.62 min (purity 97.8%). LC/MS: 464.7 (M+H)⁺.

Example 565-(2′,4′-dimethoxy-2-methylbiphenyl-4-yl)-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

μL; 1.32 mmol; 3 eq.), Intermediate 28 (120 mg; 0.44 mmol; 1 eq.),Intermediate 2 (97 mg; 0.44 mmol, 1 eq.) and DIEA (228 μL; 1.32 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 95/5) afforded the titlecompound as a white solid.

HPLC (Method A): Rt 6.11 min (purity 96.3%). LC/MS: 457.0 (M+H)⁺.

Example 573-(2-methoxyphenyl)-5-[4-(4-methyl-3-thienyl)-3-(trifluoromethyl)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (106 μL; 1.26 mmol; 3 eq.), Intermediate 32 (120 mg;0.42 mmol; 1 eq.), Intermediate 1 (70 mg; 0.42 mmol, 1 eq.) and DIEA(217 μL; 1.26 mmol; 3 eq.) were reacted according to general procedure2. Purification by column chromatography (c-hexane/ethyl acetate, 95/5)followed by crystallization from n-pentane afforded the title compoundas a yellow solid.

HPLC (Method A): Rt 5.77 min (purity 95.1%). LC/MS: 416.8 (M+H)⁺.

Example 581-(2-methyl-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)piperidine

μL; 1.64 mmol; 3 eq.), Intermediate 33 (120 mg; 0.55 mmol; 1 eq.),Intermediate 2 (120 mg; 0.55 mmol, 1 eq.) and DIEA (283 μL; 1.64 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 95/5) afforded the titlecompound as a yellow oil.

HPLC (Method A): Rt 5.62 min (purity 93.2%). LC/MS: 404.0 (M+H)⁺.

Example 591-[4-[3-(2-methoxyphenyl)-1)-1,2,4-oxadiazol-5-yl]-2-(trifluoromethyl)phenyl]piperidine

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 42 (137 mg; 0.5mmol; 1 eq.), Intermediate 1 (83 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by crystallisation from n-pentane afforded the titlecompound as a white solid.

HPLC (Method A): Rt 6.05 min (purity 99.0%). LC/MS: 404.0 (M+H)⁺.

Example 601-[4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(trifluoromethyl)phenyl]piperidine

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 42 (137 mg; 0.5mmol; 1 eq.), Intermediate 23 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by crystallisation from n-pentane afforded the titlecompound as a white solid.

HPLC (Method A): Rt 6.20 min (purity 100%). LC/MS: 422.2 (M+H)⁺.

Example 611-[4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}-2-(trifluoromethyl)phenyl]piperidine

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 42 (137 mg; 0.5mmol; 1 eq.), Intermediate 2 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by crystallisation from MeOH/water afforded the titlecompound as a white solid.

HPLC (Method A): Rt 6.74 min (purity 100%). LC/MS: 458.2 (M+H)⁺.

Example 623-(2-methoxyphenyl)-5-[2′-methyl-2-(trifluoromethyl)biphenyl-4-yl]-1,2,4-oxadiazole

Oxalyl chloride (109 μL; 1.28 mmol; 3 eq.), Intermediate 34 (120 mg;0.43 mmol; 1 eq.), Intermediate 1 (71 mg; 0.43 mmol, 1 eq.) and DIEA (95μL; 1.28 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 95/5)afforded the title compound as a white solid.

HPLC (Method A): Rt 5.86 min (purity 92.8%). LC/MS: 411.2 (M+H)⁺.

Example 631-[4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(trifluoromethyl)phenyl]-2-methylpiperidine

Oxalyl chloride (106 μL; 1.25 mmol; 3 eq.), Intermediate 24 (120 mg;0.42 mmol; 1 eq.), Intermediate 23 (77 mg; 0.42 mmol, 1 eq.) and DIEA(2164; 1.25 mmol; 3 eq.) were reacted according to general procedure 2.Purification by preparative HPLC (increasing amount of 0.1% TFA inCH₃CN, in 0.1% TFA in water) afforded the title compound as a yellowsolid.

HPLC (Method A): Rt 6.77 min (purity 96.1%). LC/MS: 436.0 (M+H)⁺.

Example 644-{4-[3-(2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylphenyl}morpholine

μL; 1.63 mmol; 3 eq.), Intermediate 35 (120 mg; 0.54 mmol; 1 eq.),Intermediate 1 (90 mg; 0.54 mmol, 1 eq.) and DIEA (280 μL; 1.63 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 95/5) afforded the titlecompound as a yellow solid.

HPLC (Method A): Rt 4.58 min (purity 99.4%). LC/MS: 352.2 (M+H)⁺.

Example 654-(2-methyl-4-{3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazol-5-yl}phenyl)morpholine

Oxalyl chloride (138 μL; 1.63 mmol; 3 eq.), Intermediate 35 (120 mg;0.54 mmol; 1 eq.), Intermediate 2 (119 mg; 0.54 mmol, 1 eq.) and DIEA(2804; 1.63 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 90/10)afforded the title compound as a white solid.

HPLC (Method A): Rt 5.53 min (purity 93.4%). LC/MS: 406.2 (M+H)⁺.

Example 664-{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylphenyl}morpholine

μL; 1.63 mmol; 3 eq.), Intermediate 35 (120 mg; 0.54 mmol; 1 eq.),Intermediate 23 (100 mg; 0.54 mmol, 1 eq.) and DIEA (280 μL; 1.63 mmol;3 eq.) were reacted according to general procedure 2. Purification byprecipitation from DCM/c-hexane afforded the title compound as a yellowsolid.

HPLC (Method A): Rt 4.78 min (purity 99.2%). LC/MS: 370.2 (M+H)⁺.

Example 671-{4-[3-(2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylphenyl}piperidine

μL; 1.64 mmol; 3 eq.), Intermediate 33 (120 mg; 0.55 mmol; 1 eq.),Intermediate 1 (91 mg; 0.55 mmol, 1 eq.) and DIEA (283 μL; 1.64 mmol; 3eq.) were reacted according to general procedure 2. Purification bypreparative HPLC (increasing amount of 0.1% TFA in CH₃CN, in 0.1% TFA inwater) afforded the title compound as an orange solid.

HPLC (Method A): Rt 4.29 min (purity 100%). LC/MS: 350.2 (M+H)⁺.

Example 681-{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylphenyl}piperidine

μL; 1.64 mmol; 3 eq.), Intermediate 33 (120 mg; 0.55 mmol; 1 eq.),Intermediate 23 (101 mg; 0.55 mmol, 1 eq.) and DIEA (283 μL; 1.64 mmol;3 eq.) were reacted according to general procedure 2. Purification bypreparative HPLC (increasing amount of 0.1% TFA in CH₃CN, in 0.1% TFA inwater) afforded the title compound as a white solid.

HPLC (Method A): Rt 4.64 min (purity 100%). LC/MS: 368.2 (M+H)⁺.

Example 695-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(2-methylpiperidin-1-yl)aniline

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 8 (132 mg; 0.5mmol; 1 eq.), Intermediate 23 (92 mg; 0.5 mmol; 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2. Afterpurification by column chromatography (c-hexane/ethyl acetate, 80/20),the residue was taken up in EtOH (20 mL) and stannous chloride dihydrate(564 mg; 2.5 mmol; 5 eq.) was added. The resulting mixture was stirredat reflux for 3 hours then concentrated in vacuo. The residue waspartitioned between NaOH 0.05M and ethyl acetate. The phases wereseparated and the organic layer was washed with brine, dried overmagnesium and concentrated in vacuo. Purification by columnchromatography (c-hexane/ethyl acetate, 75/25) afforded the titlecompound as an off-white solid.

HPLC (Method A): Rt 3.50 min (purity 97.7%). LC/MS: 383.3 (M+H)⁺.

Example 705-[4-(4-methyl-3-thienyl)-3-(trifluoromethyl)phenyl]-3-[2-(trifluoromethoxy)phenyl]-1,2,4-oxadiazole

Oxalyl chloride (106 μL; 1.26 mmol; 3 eq.), Intermediate 32 (120 mg;0.42 mmol; 1 eq.), Intermediate 2 (92 mg; 0.42 mmol, 1 eq.) and DIEA(217 μL; 1.26 mmol; 3 eq.) were reacted according to general procedure2. Purification by crystallisation from MeOH afforded the title compoundas a white solid.

HPLC (Method A): Rt 6.57 min (purity 98.3%). LC/MS: 470.4 (M+H)⁺.

Example 715-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(2-methylpiperidin-1-yl)benzonitrile

Oxalyl chloride (190 mg; 1.5 mmol; 3 eq.), Intermediate 36 (122 mg; 0.5mmol; 1 eq.), Intermediate 23 (92 mg; 0.5 mmol, 1 eq.) and DIEA (194 mg;1.5 mmol; 3 eq.) were reacted according to general procedure 2.Purification by column chromatography (c-hexane/ethyl acetate, 70/30)afforded the title compound as a white solid.

HPLC (Method A): Rt 5.47 min (purity 96.7%). LC/MS: 393.2 (M+H)⁺.

Example 72N-[5-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(2-methylpiperidin-1-yl)phenyl]methanesulfonamide

(83 mg; 0.22 mmol; 1 eq.) in pyridine (1 mL) and the resulting solutionwas stirred at room temperature for 16 hours. The solution was thenpartitioned between water and ethyl acetate. The two phases wereseparated and the aqueous layer was extracted with ethyl acetate. Thecombined organic phase was washed (3×) with 0.1 M HCl, dried overmagnesium sulfate and concentrated in vacuo. Purification by columnchromatography (c-hexane/ethyl acetate, 60/40) afforded the titlecompound as a white solid.

HPLC (Method A): Rt 4.93 min (purity 100%). LC/MS: 461.2 (M+H)⁺.

Example 733-{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylphenyl}-2-methylpyridine

te 37 (50 mg; 0.22 mmol; 1 eq.), Intermediate 23 (41 mg; 0.22 mmol, 1eq.) and DIEA (85 mg; 0.66 mmol; 3 eq.) were reacted according togeneral procedure 2. Purification by column chromatography(c-hexane/ethyl acetate, 90/10 then 85/15) afforded the title compoundas a white solid.

HPLC (Method A): Rt 3.19 min (purity 94.2%). LC/MS: 376.3 (M+H)⁺.

Example 745-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-3-methyl-2-(2-methylpiperidin-1-yl)pyridine

μL; 1.54 mmol; 3 eq.), Intermediate 38 (120 mg; 0.51 mmol; 1 eq.),Intermediate 23 (94 mg; 0.51 mmol, 1 eq.) and DIEA (260 μL; 1.54 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 50/50) afforded the titlecompound as a white solid.

HPLC (Method A): Rt 4.38 min (purity 88.7%). LC/MS: 383.3 (M+H)⁺.

Example 755-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-(2-methylpiperidin-1-yl)pyridine

Oxalyl chloride (138 μL; 1.63 mmol; 3 eq.), Intermediate 39 (120 mg;0.54 mmol; 1 eq.), Intermediate 23 (100 mg; 0.54 mmol, 1 eq.) and DIEA(282 μL; 1.54 mmol; 3 eq.) were reacted according to general procedure2. Purification by column chromatography (c-hexane/ethyl acetate, 50/50)afforded the title compound as a yellow oil.

HPLC (Method A): Rt 3.92 min (purity 84.3%). LC/MS: 369.3 (M+H)⁺.

Example 765-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-[2-(methoxymethyl)pyrrolidin-1-yl]-3-methylpyridine

μL; 1.44 mmol; 3 eq.), Intermediate 40 (120 mg; 0.48 mmol; 1 eq.),Intermediate 23 (88 mg; 0.48 mmol, 1 eq.) and DIEA (245 μL; 1.44 mmol; 3eq.) were reacted according to general procedure 2. Purification bycolumn chromatography (c-hexane/ethyl acetate, 50/50) afforded the titlecompound as a yellow oil.

HPLC (Method A): Rt 3.49 min (purity 95.8%). LC/MS: 399.3 (M+H)⁺.

Example 77{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2′-methylbiphenyl-2-yl}methanol

Intermediate 23 (276 mg; 1.5 mmol; 1 eq.), Intermediate 41 (436 mg; 1.8mmol; 1.2 eq.) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride (345 mg; 1.8 mmol; 1.2 eq.) were dissolved in THF (6 mL)and acetonitrile (6 mL) and the reaction mixture was stirred at roomtemperature for 2 hours. DIEA (0.61 mL; 3.6 mmol; 2.4 eq.) was added andthe mixture was heated in the microwave at 150° C. for 30 min. Thereaction mixture was then filtered through a SPE-NH₂ column (2 g) andthrough a SPE-SCX column (2 g) and rinsed with ACN. The filtrate wasevaporated and the residue washed with ACN to afford the title compoundas a white solid.

HPLC (Method A), Rt: 5.00 min (purity: 98.8%). LC/MS: 391.1 (M+H)⁺. ¹HNMR (CDCl₃, 300 MHz) δ 8.45 (d, J=1.3 Hz, 1H), 8.18 (dd, J=7.9, 1.8 Hz,1H), 7.91 (dd, J=9.0, 3.2 Hz, 1H), 7.35-7.13 (m, 6H), 7.03 (dd, J=9.2,4.3 Hz, 1H), 4.51 (s, 2H), 4.00 (s, 3H), 2.08 (s, 3H), 1.61 (br s, 1H).

Example 781-{4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2′-methylbiphenyl-2-yl}-N,N-dimethylmethanamine

To a solution of Example 77 (115 mg; 0.29 mmol; 1 eq.) in DCM (6 mL) isadded at 0° C. N-ethyldiisopropylamine (250 μL; 1.47 mmol; 5 eq.) andmethanesulfonyl chloride (27 μL; 0.35 mmol; 1.2 eq.). After 15 min,dimethylamine (2M; 440 μL; 0.88 mmol; 3 eq.) was added and the reactionmixture stirred at room temperature for 2 hours. Water (5 mL) was addedand the organic layer washed with water. The organic phase was driedover magnesium sulfate and concentrated in vacuo. The residue waspurified by mass triggered preparative HPLC (increasing amount of ACN inwater) to afford the title compound as a brown oil.

HPLC (Method A), Rt: 3.81 min (purity: 98.9%). LC/MS: 418.2 (M+H)⁺. ¹HNMR (DMSO-d₆, 300 Hz) δ 8.37 (s, 1H), 8.01 (d, J=8.0 Hz, 1H), 7.77 (d,J=7.8 Hz, 1H), 7.50-7.45 (m, 1H), 7.39-7.29 (m, 5H), 7.11 (d, J=6.8 Hz,1H), 3.91 (s, 3H), 3.18 (s, 2H), 2.07 (s, 6H), 2.01 (s, 3H).

Example 794-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2′-methylbiphenyl-2-carboxylicacid

Step 14-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2′-methylbiphenyl-2-carbaldehyde

Manganese(IV) oxide (111 mg; 1.28 mmol; 10 eq.) was added to a solutionof Example 77 (50 mg; 0.13 mmol; 1 eq.) in DCM (5 mL) and the reactionmixture was stirred at room temperature for 4 hours. The suspension wasfiltered through a pad of Celite® and the solution concentrated in vacuoto afford title compound (46 mg; 93%) as a white solid.

HPLC (Method A), Rt: 5.48 min (purity: 98.7%). LC/MS: 389.2 (M+H)⁺.

Step 2:4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2′-methylbiphenyl-2-carboxylicacid

A solution of sodium chlorite (59 mg; 0.65 mmol; 5.5 eq.) and sodiumdihydrogenphosphate (51 mg; 0.43 mmol; 3.6 eq.) in water (1 mL) wasadded to a mixture of 2-methyl-2-butene (0.13 mL) and4-[3-(5-fluoro-2-methoxyphenyl)-1,2,4-oxadiazol-5-yl]-2-methylbiphenyl-2-carbaldehyde(46.2 mg; 0.12 mmol; 1 eq.) in dioxane (1 mL) and the reaction wasstirred at room temperature for 16 hours. The suspension was partitionedbetween water and ethyl acetate. The aqueous phase was acidified to pH3-4 with acetic acid and extracted with ethyl acetate. The combinedorganic phase was washed with brine, dried over magnesium sulfate andconcentrated in vacuo to afford the title compound (43 mg; 89%) as awhite solid.

HPLC (Method A), Rt: 5.19 min (purity: 99.7%). LC/MS: 405.2 (M+H)⁺. ¹HNMR (DMSO-d₅, 300 MHz) δ 13.09 (br s, 1H), 8.59 (d, J=1.8 Hz, 1H), 8.34(dd, J=8.0, 1.9 Hz, 1H), 7.80 (dd, J=9.0, 3.2 Hz, 1H), 7.53 (d, J=8.0Hz, 1H), 7.50-7.44 (m, 1H), 7.33-7.21 (m, 4H), 7.10 (d, J=7.0 Hz, 1H),3.92 (s, 3H), 2.07 (s, 3H).

Example 80 In Vitro Assays

Receptor binding assay: Membranes were prepared from CHO cellsexpressing S1P₁ or S1P₃ for use in ligand and 35S-GTPγS binding studies.Cells were suspended in 50 mM TRIS, pH 7.4, 2 mM EDTA, 250 mM Sucrose(buffer A) and 1× Complete protease inhibitor cocktail (Roche), anddisrupted at 4° C. by nitrogen decompression using a cell disruptionbomb (Parr Instrument). Following centrifugation at 1000 RPM for 10 minat 4° C., the supernatant was suspended in buffer A and centrifugedagain at 19000 RPM for 60 min at 4° C. The pellet was then suspended in10 mM HEPES, pH 7.4, 1 mM EDTA, 250 mM Sucrose (Buffer B), and 1×Complete EDTA-free protease inhibitor cocktail and homogenized using apotter. Membranes were flash frozen in liquid nitrogen and stored at−80° C. [33P]sphingosine 1-phosphate (3000 Ci/mmol; AmericanRadiolabeled Chemicals, Inc.) was added to test compounds in DMSO.Membranes and WGA SPA beads (GE Healthcare) were added to give a finalvolume of 100 μl in 96-well plates with assay concentrations of 25 pM or10 pM [33P]sphingosine 1-phosphate (respectively for S1P1 or S1P3), 50mM HEPES, pH 7.5, 5 mM MgCl2, 100 mM NaCl, 0.4% fatty acid-free BSA, 1-5μg/well of proteins and 100 μg/well of WGA SPA beads. Binding wasperformed for 60 min at room temperature on a shaker and boundradioactivity was measured on a PerkinElmer 1450 MicroBeta counter.Specific binding was calculated by subtracting remaining radioactivityin the presence of 1000-fold excess of unlabeled S1P. Binding data wereanalyzed using the GraphPad Prism program.

Measurements of ³⁵S-GTPγS Binding: Membranes (1 to 10 pg protein)prepared as described above, were incubated in 96-well Scintiplates(Perkin Elmer) with test compounds diluted in DMSO, in 180 μl of 20 mMHEPES, pH 7.4, 10 mM MgCl₂, 2 μg/well Saponin, 0.2% fatty acid free BSA(Assay buffer), 140 mM NaCl and 1.7 μM GDP. The assay was initiated withthe addition of 20 μl of 1.5 nM [35S]-GTPγS (1100 Ci/mmol; GEHealthcare) in assay buffer. After 60 min incubation at 30° C. on ashaker, plates were centrifuged for 10 min at 2000 RPM. Supernatant wasdiscarded and membrane bound radioactivity was measured on a PerkinElmer1450 MicroBeta counter. Triplicate samples were averaged and expressedas % response relative to S1P activation in absence of compound (n=2).

The compound of formula (I) have utility as immunoregulatory agents asdemonstrated by their activity as potent and selective agonists of theS1P₁ receptor over the S1P₃ receptor as measured in the assays describedabove. In particular, the examples disclosed herein possess aselectivity for the S1P1 receptor over the S1P3 receptor as measured bythe ratio of EC50 for the S1P1 receptor to the EC50 for the S1P₃receptor as evaluated in the ³⁵S-GTPγS binding assay described above.The following results have been obtained:

S1P₁ S1P₁ S1P₃ Binding GTPgS GTPgs Compound Ki (μM) EC50 (μM) EC50 (μM)I1 

0.011 0.027 >20 I2 

0.018 0.051 >30 I3 

0.652 1.785 — I4 

1.074 3.605 — I5 

— 0.589 — I6 

— 0.712 — I7 

— 0.254 — I8 

— 0.018 >30 I9 

— 0.007 5.410 I10

— 0.024 >30 I11

— 0.080 — I12

— 0.036 >30 I13

— 0.132 — I14

— 0.428 — I15

— 1.130 — I16

— 0.228 — I17

— 0.135 — I18

0.007 0.006 >30 I19

— 0.018 >30 I20

0.016 0.044 3.210 I21

0.008 0.004 >30 I22

— 0.051 >30 I23

0.016 0.010 >30 I24

— 0.057 — I25

0.003 0.009 5.485 I26

0.002 0.009 5.050 I27

0.020 0.066 >30 I28

0.091 0.162 >30 I29

0.025 0.059 — I30

— 0.064 — I31

— 0.058 — I32

0.001 0.002 1.705 I33

0.043 0.006 — I34

0.005 0.006 >20 I35

0.003 0.002 0.650 I36

— 0.010 2.100 I37

— 0.921 — I38

— 4.470 — I39

— 0.267 — I40

— 0.027 — I41

— 0.116 — I42

— 1.251 — I43

— 0.039 — I44

0.005 0.003 >20 I45

— 0.010 >20 I46

— 0.033 — I47

— 1.200 — I48

0.004 0.005 2.180 I49

— 0.191 — I50

— 0.172 — I51

— 0.085 — I52

— 0.295 — I53

— 0.014 >20 I54

— 0.013 >30 I55

0.008 0.002 >30 I56

— 0.302 — I57

0.002 0.001 >20 I58

— 0.039 — I59

— 0.021 — I60

0.001 0.002 0.617 I61

0.054 0.004 — I62

0.008 0.004 — I63

0.003 0.001 0.125 I64

— 0.098 — I65

— 0.130 — I66

— 0.028 — I67

— 0.024 — I68

— 0.010 — I69

— 0.024 0.270 I70

0.004 0.002 — I71

0.004 0.001 0.074 I72

0.046 0.008 >20 I73

— 0.341 — I74

0.006 0.005 — I75

— 1.185 — I76

— 0.522 — I77

0.053 0.065 5.61  I78

0.004 0.002 0.092 I79

— 0.657 —

Example 32 Animal Models Evaluating the In Vivo Efficacy of S1P AgonistsModel of S1P Agonists-Induced Lymphopenia in Mice

Female C57BL/6 mice (Elevage Janvier) (8 week old) receive S1P agonistsby oral route. Blood is sampled in heparinized (100 IU/kg, ip) mice byintracardiac or retroorbital puncture under isoflurane anesthesia 2 to120 hrs after drug treatment. The white blood cells (lymphocytes andneutrophils) are counted using a Beckman/Coulter counter. The quality ofblood sampling is assessed by counting erythocytes and platelets.

Model of MOG-Induced Experimental Autoimmune Encephalomyelytis (Eae) inMice

EAE was induced in 9 weeks old female mice (C57BL/6, Elevage Janvier) byan immunization against MOG. The mice received Pertussis toxin (Alexis,300 ng/mouse in 200 μl of PBS) by ip route and 100 μl of an emulsioncontaining MOG35-55 peptide (NeoMPS, 200 μg/mouse), MycobacteriumTuberculosis (0.25 mg/mouse) in Complete Freund's Adjuvant (DIFCO) bysubcutaneous injection into the back. Two days later an additionalinjection of Pertussis toxin (Alexis, 300 ng/mouse in 200 μl of PBS) wasdone by ip route. After EAE induction, mice were weighed daily and theneurological impairment was quantified using a 15-points clinical scaleassessing the paralysis (tail, hind limbs and fore limbs), theincontinency and the death.

Clinical Score

—1— Tail

-   -   Score=0 A normal mouse holds its tail erect when moving.    -   Score=1 If the extremity of the tail is flaccid with a tendency        to fall.    -   Score=2 If the tail is completely flaccid and drags on the        table.

—2— Hind Limbs

-   -   Score=0 A normal mouse has an energetic walk and doesn't drag        his paws.    -   Score=1 Either one of the following tests is positive:        —a— Flip test: while holding the tail between thumb and index        finger, flip the animal on his back and observe the time it        takes to right itself. A healthy mouse will turn itself        immediately. A delay suggests hind-limb weakness.        —b— Place the mouse on the wire cage top and observe as it        crosses from one side to the other. If one or both limbs        frequently slip between the bars we consider that there is a        partial paralysis.    -   Score=2 Both previous tests are positive.    -   Score=3 One or both hind limbs show signs of paralysis but some        movements are preserved; for example: the animal can grasp and        hold on to the underside of the wire cage top for a short moment        before letting go    -   Score=4 When both hind legs are paralyzed and the mouse drags        them when moving.

—3— Fore Limbs:

-   -   Score=0 A normal mouse uses his front paws actively for grasping        and walking and holds his head erect.    -   Score=1 Walking is possible but difficult due to a weakness in        one or both of the paws, for example, the front paws are        considered weak when the mouse has difficulty grasping the        underside of the wire top cage. Another sign of weakness is head        drooping.    -   Score=2 When one forelimb is paralyzed (impossibility to grasp        and the mouse turns around the paralyzed limb). At this time the        head has also lost much of its muscle tone.    -   Score=3 Mouse cannot move, and food and water are unattainable.

—4— Bladder:

Score=0 A normal mouse has full control of his bladder.Score=1 A mouse is considered incontinent when his lower body is soakedwith urine.

—5— Death:

-   -   Score=15        The final score for each animal is determined by the addition of        all the above-mentioned categories. The maximum score for live        animals is 10.

At day 12 (first signs of paralysis) the mice were stratified inexperimental groups (n=10) according to the clinical score and the bodyweight loss. The semi-curative treatment started at day 14.

Example A Injection Vials

A solution of 100 g of an active ingredient of the formula I and 5 g ofdisodium hydrogenphosphate in 3 L of bidistilled water is adjusted to pH6.5 using 2 N hydrochloric acid, sterile filtered, transferred intoinjection vials, lyophilised under sterile conditions and sealed understerile conditions. Each injection vial contains 5 mg of activeingredient.

Example B Suppositories

A mixture of 20 g of an active ingredient of the formula I with 100 g ofsoya lecithin and 1400 g of cocoa butter is melted, poured into mouldsand allowed to cool. Each suppository contains 20 mg of activeingredient.

Example C Solution

A solution is prepared from 1 g of an active ingredient of the formula(I), 9.38 g of NaH₂PO₄.2H₂O, 28.48 g of Na₂HPO₄-12H₂O and 0.1 g ofbenzalkonium chloride in 940 mL of bidistilled water. The pH is adjustedto 6.8, and the solution is made up to 1 L and sterilised byirradiation. This solution can be used in the form of eye drops.

Example D Ointment

500 mg of an active ingredient of the formula (I) are mixed with 99.5 gof Vaseline under aseptic conditions.

Example E Tablets

A mixture of 1 kg of active ingredient of the formula I, 4 kg oflactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesiumstearate is pressed to give tablets in a conventional manner in such away that each tablet contains 10 mg of active ingredient.

Example F Coated Tablets

Tablets are pressed analogously to Example E and subsequently coated ina conventional manner with a coating of sucrose, potato starch, talc,tragacanth and dye.

Example G Capsules

2 kg of active ingredient of the formula I are introduced into hardgelatine capsules in a conventional manner in such a way that eachcapsule contains 20 mg of the active ingredient.

Example H Ampoules

A solution of 1 kg of active ingredient of the formula I in 60 L ofbidistilled water is sterile filtered, transferred into ampoules,lyophilised under sterile conditions and sealed under sterileconditions. Each ampoule contains 10 mg of active ingredient.

1-15. (canceled)
 16. A compound of Formula I

wherein: R¹, R² denote H, Hal, CF₃, OCF₃, CN, or NO₂, W denotes CH or N,R^(a) is Ar, Het, cycloalkyl having 3-7 atoms, A or NA₂, R^(b) is H, A,Hal, CF₃, OCF₃, OR³, CN, NO₂, (CH₂)_(n)N(R³)₂, OA, (CH₂)_(n)SO₂N(R³)₂,(CH₂)_(n)NR³SO₂A, (CH₂)_(n)N(SO₂A)₂, NR³CON(R³)₂, NR³COA or(CH₂)_(n)SO₂R³, R^(e) denotes A, COA, CSA, COOA, CSOA, CON(R³)₂ orCSN(R³)₂, A is branched or linear alkyl having 1 to 12 C-atoms, whereinone or more, H-atoms may be replaced by Hal, or 1 to 7H-atoms may bereplaced by OR³, CN or N(R³)₂ and wherein one or more, preferably 1 to 7non-adjacent CH₂-groups may be replaced by O, NR³ or S and/or by —CH═CH—or —C≡C— groups, or denotes cycloalkyl or cycloalkylalkylene having 3-7ring C atoms, Hal is F, Cl, Br or I, Ar denotes a monocyclic orbicyclic, unsaturated or aromatic carbocyclic ring having 6 to 14 carbonatoms which may be unsubstituted or monosubstituted, disubstituted ortrisubstituted by Hal, A, CH₂OA, CH₂N(R³)₂, OR³, N(R³)₂, NO₂, N(SO₂Me)₂,CN, COOR³, CF₃, OCF₃, CON(R³)₂, NR³COA, NR³CON(R³)₂, NR³SO₂A, COR³,SO₂N(R³)₂, SOA or SO₂A, phenyl, pyridyl-[C(R³)₂]_(n)—COOR³ and/or—O[C(R³)₂]_(n)—CON(R³)₂, Het denotes a monocyclic or bicyclic,saturated, unsaturated or aromatic heterocyclic ring having 1 N and/or 1to 3O and/or S atoms which may be unsubstituted or monosubstituted,disubstituted or trisubstituted by Hal, A, —[C(R³)₂]_(n)—Ar,—[C(R³)₂]_(n)-cycloalkyl, CH₂OA, CH₂N(R³)₂, OR³, CF₃, OCF₃, N(R³)₂,N(SO₂Me)₂, NR³CON(R³)₂, NO₂, CN, —[C(R³)₂]_(n)—COOR³,—[C(R³)₂]_(n)—CON(R³)₂, NR³COA, NR³SO₂A, COR³, SO₂N(R³)₂, SOA, phenyl,pyridyl and/or SO₂A, R³ is H or A, and n is 0, 1, 2, 3, 4, 5, 6, 7 or 8,and pharmaceutically acceptable solvates, tautomers, salts andstereoisomers thereof, including mixtures thereof in all ratios.
 17. Thecompound according to claim 16, wherein R^(a) denotes Ar or Het.
 18. Thecompound according to claim 17, wherein Ar denotes one of the followinggroups:

wherein X¹, and X² independently of one another F, OCH₃, CH₃, CF₃, OCF₃,OH, NO₂, CN, and/or phenyl.
 19. The compound according to claim 17,wherein Het denotes one of the following groups:

wherein X¹, and X² denote independently of one another F, OCH₃, CH₃,CF₃, OCF₃, OH, NO₂, CN, and/or phenyl and R³ has the meaning given inclaim
 16. 20. The compound according to claim 17, wherein Ar or Het issubstituted by methyl, trifluoromethyl or methoxy.
 21. The compoundaccording to claim 16, said compound being selected from:

and pharmaceutically acceptable solvates, tautomers, salts andstereoisomers thereof, including mixtures thereof in all ratios.
 22. Thecompound according to claim 16, wherein said compound has an EC50 inGTPγS for the binding to the S1P₁ receptor of less than about 5 μM. 23.A process for the preparation of a compound according to claim 16comprising reacting: a) a compound of formula A

wherein W, R, R^(a) and R^(b) have the meanings given above, and T isOH, or a leaving group, or the product of the reaction of isobutylchloroformate with formula A, wherein T is OH; and b) a compound offormula B

wherein R¹, R² and R^(c) have the meanings given in claim 16 in thepresence of a base, or in case T is OH, in the presence of a suitablecondensation reagent, and cyclizing the product, and optionallyconverting the compound into a salt.
 24. A pharmaceutical compositioncomprising at least one compound according to claim 16 and an excipientor adjuvant.
 25. The pharmaceutical composition according to claim 24,further comprising at least one further active ingredient.
 26. A kitconsisting of separate packs of: (a) an effective amount of a compoundaccording to claim 16, and (b) an effective amount of an additionalactive ingredient.
 27. A method for the treatment and/or prophylaxis ofa sphingosine 1-phosphate associated disorder comprising theadministration of a composition according to claim 24 to an individualhaving said disorder.
 28. The method according to claim 27, wherein thesphingosine 1-phosphate-(1) associated disorder is an autoimmunedisorder or condition associated with an overactive immune response. 29.The method according to claim 27, wherein said disorder is animmuneregulatory abnormality.
 30. The method according to claim 29,wherein the immunoregulatory abnormality is an autoimmune or chronicinflammatory disease selected from the group consisting of: systemiclupus erythematosis, chronic rheumatoid arthritis, type I diabetesmellitus, inflammatory bowel disease, biliary cirrhosis, uveitis,multiple sclerosis, amyotrophic lateral sclerosis (ALS), Crohn'sdisease, ulcerative colitis, bullous pemphigoid, sarcoidosis, psoriasis,autoimmune myositis, Wegener's granulomatosis, ichthyosis, Gravesopthalmopathy, bone marrow, organ transplant rejection orgraft-versus-host disease, transplantation of organs or tissue,graft-versus-host diseases brought about by transplantation, autoimmunesyndromes including, rheumatoid arthritis, Hashimoto's thyroiditis,myasthenia gravis, type I diabetes, uveitis, posterior uveitis, allergicencephalomyelitis, glomerulonephritis, post-infectious autoimmunediseases including rheumatic fever and post-infectiousglomerulonephritis, inflammatory and hyperproliferative skin diseases,psoriasis, atopic dermatitis, contact dermatitis, eczematous dermatitis,seborrhoeic dermatitis, lichen planus, pemphigus, bullous pemphigoid,epidermolysis bullosa, urticaria, angioedemas, vasculitis, erythema,cutaneous eosinophilia, lupus erythematosus, acne, alopecia greata,keratoconjunctivitis, vernal conjunctivitis, uveitis associated withBehcet's disease, keratitis, herpetic keratitis, conical cornea,dystrophia epithelialis corneae, corneal leukoma, ocular pemphigus,Mooren's ulcer, scleritis, Graves' opthalmopathy, Vogt-Koyanagi-Haradasyndrome, sarcoidosis, pollen allergies, reversible obstructive airwaydisease, bronchial asthma, allergic asthma, intrinsic asthma, extrinsicasthma, dust asthma, chronic or inveterate asthma, late asthma andairway hyper-responsiveness, bronchitis, gastric ulcers, vascular damagecaused by ischemic diseases and thrombosis, ischemic bowel diseases,inflammatory bowel diseases, necrotizing enterocolitis, intestinallesions associated with thermal burns, coeliac diseases, proctitis,eosinophilic gastroenteritis, mastocytosis, migraine, rhinitis, eczema,interstitial nephritis, Goodpasture's syndrome, hemolytic-uremicsyndrome, diabetic nephropathy, multiple myositis, Guillain-Barresyndrome, Meniere's disease, polyneuritis, multiple neuritis,mononeuritis, radiculopathy, hyperthyroidism, Basedow's disease, purered cell aplasia, aplastic anemia, hypoplastic anemia, idiopathicthrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis,pernicious anemia, megaloblastic anemia, anerythroplasia, osteoporosis,sarcoidosis, fibroid lung, idiopathic interstitial pneumonia,dermatomyositis, leukoderma vulgaris, ichthyosis vulgaris, photoallergicsensitivity, cutaneous T cell lymphoma, chronic lymphocytic leukemia,arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritisnodosa, myocardosis, scleroderma, Wegener's granuloma, Sjogren'ssyndrome, adiposis, eosinophilic fascitis, lesions of gingiva,periodontium, alveolar bone, substantia ossea dentis,glomerulonephritis, male pattern alopecia or alopecia senilis bypreventing epilation or providing hair germination and/or promoting hairgeneration and hair growth, muscular dystrophy, pyoderma and Sezary'ssyndrome, Addison's disease, ischemia-reperfusion injury of organs whichoccurs upon preservation, transplantation or ischemic disease,endotoxin-shock, pseudomembranous colitis, colitis caused by drug orradiation, ischemic acute renal insufficiency, chronic renalinsufficiency, toxinosis caused by lung-oxygen or drugs, lung cancer,pulmonary emphysema, cataracta, siderosis, retinitis pigmentosa, senilemacular degeneration, vitreal scarring, corneal alkali burn, dermatitiserythema multiforme, linear IgA ballous dermatitis and cementdermatitis, gingivitis, periodontitis, sepsis, pancreatitis, diseasescaused by environmental pollution, aging, carcinogenesis, metastasis ofcarcinoma and hypobaropathy, disease caused by histamine orleukotriene-C₄ release, Behcet's disease, autoimmune hepatitis, primarybiliary cirrhosis, sclerosing cholangitis, partial liver resection,acute liver necrosis, necrosis caused by toxin, viral hepatitis, shock,or anoxia, B-virus hepatitis, non-A/non-B hepatitis, cirrhosis,alcoholic cirrhosis, hepatic failure, fulminant hepatic failure,late-onset hepatic failure, “acute-on-chronic”liver failure,augmentation of chemotherapeutic effect, cytomegalovirus infection, HCMVinfection, AIDS, cancer, senile dementia, trauma, and chronic bacterialinfection and asthma.